bims-auttor 2020-01-12 papers

bims-auttor

Biomed News

on Autophagy and mTOR

Issue of 2020–01–12
thirty-one papers selected by
Viktor Korolchuk, Newcastle University

NBR1-mediated p62-liquid droplets enhance the Keap1-Nrf2 system.
Age-dependent autophagy induction after injury promotes axon regeneration by limiting NOTCH.
Control of Inflammation by Calorie Restriction Mimetics: On the Crossroad of Autophagy and Mitochondria.
The GATOR2-mTORC2 axis mediates Sestrin2-induced AKT Ser/Thr kinase activation.
mTORC1-Rps15 Axis Contributes to the Mechanisms Underlying Global Translation Reduction During Senescence of Mouse Embryonic Fibroblasts.
The ATG conjugation systems in autophagy.
Exploring Molecular Descriptors and Fingerprints to Predict mTOR Kinase Inhibitors using Machine Learning Techniques.
Cell-type specific regulation of neuronal intrinsic excitability by macroautophagy.
UVRAG in autophagy, inflammation, and cancer.
Fas-apoptotic inhibitory molecule 2 localizes to the lysosome and facilitates autophagosome-lysosome fusion through the LC3 interaction region motif-dependent interaction with LC3.
Innate immune receptor NOD2 mediates LGR5+ intestinal stem cell protection against ROS cytotoxicity via mitophagy stimulation.
Mitophagy and DNA Damage Signaling in Human Aging.
Beclin 1 regulates recycling endosome and is required for skin development in mice.
TRIM14 Promotes Noncanonical NF-κB Activation by Modulating p100/p52 Stability via Selective Autophagy.
p53 regulates mitochondrial dynamics by inhibiting Drp1 translocation into mitochondria during cellular senescence.
Pathogenic Pathways in Early-Onset Autosomal Recessive Parkinson's Disease Discovered Using Isogenic Human Dopaminergic Neurons.
Autophagy, Cellular Aging and Age-related Human Diseases.
Promoting tau secretion and propagation by hyperactive p300/CBP via autophagy-lysosomal pathway in tauopathy.
mTORC1 as a Regulator of Mitochondrial Functions and a Therapeutic Target in Cancer.
SQSTM1/p62 activates NFE2L2/NRF2 via ULK1-mediated autophagic KEAP1 degradation and protects mouse liver from lipotoxicity.
Use of the LC3B-fusion technique for biochemical and structural studies of proteins involved in the N-degron pathway.
The Sigma-1 Receptor at the Crossroad of Proteostasis, Neurodegeneration, and Autophagy.
A dual druggable genome-wide siRNA and compound library screening approach identifies modulators of parkin recruitment to mitochondria.
Autophagy-Dependent Ferroptosis Drives Tumor-Associated Macrophage Polarization via Release and Uptake of Oncogenic KRAS Protein.
Inhibition of the glutamine transporter SNAT1 confers neuroprotection in mice by modulating the mTOR-autophagy system.
Transcriptional regulation of autophagy-lysosomal pathway in cancer.
Autophagy and LRRK2 in the Aging Brain.
Autophagy mediates hepatic GRK2 degradation to facilitate glucagon-induced metabolic adaptation to fasting.
ULK1 phosphorylates Exo70 to suppress breast cancer metastasis.
UPRER promotes lipophagy independent of chaperones to extend life span.
Loss of TSC complex enhances gluconeogenesis via upregulation of Dlk1-Dio3 locus miRNAs.

EMBO Rep. 2020 Jan 09. e48902

NBR1-mediated p62-liquid droplets enhance the Keap1-Nrf2 system.

Pablo Sánchez-Martín, Yu-Shin Sou, Shun Kageyama, Masato Koike, Satoshi Waguri, Masaaki Komatsu.

  p62/SQSTM1 is a multivalent protein that has the ability to cause liquid-liquid phase separation and serves as a receptor protein that participates in cargo isolation during selective autophagy. This protein is also involved in the non-canonical activation of the Keap1-Nrf2 system, a major oxidative stress response pathway. Here, we show a role of neighbor of BRCA1 gene 1 (NBR1), an autophagy receptor structurally similar to p62/SQSTM1, in p62-liquid droplet formation and Keap1-Nrf2 pathway activation. Overexpression of NBR1 blocks selective degradation of p62/SQSTM1 through autophagy and promotes the accumulation and phosphorylation of p62/SQSTM1 in liquid-like bodies, which is required for the activation of Nrf2. NBR1 is induced in response to oxidative stress, which triggers p62-mediated Nrf2 activation. Conversely, loss of Nbr1 suppresses not only the formation of p62/SQSTM1-liquid droplets, but also of p62-dependent Nrf2 activation during oxidative stress. Taken together, our results show that NBR1 mediates p62/SQSTM1-liquid droplet formation to activate the Keap1-Nrf2 pathway.

Keywords:  NBR1; Nrf2; autophagy; liquid droplet; p62/SQSTM1

DOI:  https://doi.org/10.15252/embr.201948902
Autophagy. 2020 Jan 10.

Age-dependent autophagy induction after injury promotes axon regeneration by limiting NOTCH.

Su-Hyuk Ko, Ellen C Apple, Zhijie Liu, Lizhen Chen.

  Macroautophagy/autophagy is essential for maintaining cellular homeostasis through the degradation of organelles and proteins. It also has a prominent role in modulating aging. However, the role of autophagy in the neuronal response to axon injury and axon regeneration, particularly in the context of aging, remains largely unknown. Our candidate genetic screen for axon regeneration regulators has identified genes in the autophagy pathway. Using a reporter that monitors autophagosomes and autolysosomes, we were able to monitor the dynamics of autophagy during axon regeneration. In response to axon injury, there was a significant increase in the number of autophagic vesicles. Injury-triggered autophagy activation and axon regeneration capacity undergo an age-dependent decline, and autophagy-activating agents partially rescued these declines. We found that DLK-1 was both required and sufficient for injury-induced autophagy activation. Autophagic vesicles co-localized with the NOTCH4 ortholog, LIN-12 receptor, a previously identified inhibitor of axon regeneration. Epistasis analyses indicate that LIN-12 might be a target of autophagy in axon regeneration. Together, our data suggest that DLK-mediated injury signaling can activate autophagy, which might limit the level of LIN-12 and NOTCH proteins to promote axon regeneration. Our findings reveal that autophagy activation can promote axon regeneration in neurons that lack maximal regrowth capacity, providing a promising therapeutic strategy for axon injury.

Keywords:  Aging; DLK; LC3; Notch signaling; autophagy; axon injury; axon regeneration

DOI:  https://doi.org/10.1080/15548627.2020.1713645
Cells. 2019 Dec 28. pii: E82. [Epub ahead of print]9(1):

Control of Inflammation by Calorie Restriction Mimetics: On the Crossroad of Autophagy and Mitochondria.

Enrique Gabandé-Rodríguez, Manuel M Gómez de Las Heras, María Mittelbrunn.

  Mitochondrial metabolism and autophagy are two of the most metabolically active cellular processes, playing a crucial role in regulating organism longevity. In fact, both mitochondrial dysfunction or autophagy decline compromise cellular homeostasis and induce inflammation. Calorie restriction (CR) is the oldest strategy known to promote healthspan, and a plethora of CR mimetics have been used to emulate its beneficial effects. Herein, we discuss how CR and CR mimetics, by modulating mitochondrial metabolism or autophagic flux, prevent inflammatory processes, protect the intestinal barrier function, and dampen both inflammaging and neuroinflammation. We outline the effects of some compounds classically known as modulators of autophagy and mitochondrial function, such as NAD+ precursors, metformin, spermidine, rapamycin, and resveratrol, on the control of the inflammatory cascade and how these anti-inflammatory properties could be involved in their ability to increase resilience to age-associated diseases.

Keywords:  aging; autophagy; calorie restriction; inflammation; metabolism; mitochondria

DOI:  https://doi.org/10.3390/cells9010082
J Biol Chem. 2020 Jan 08. pii: jbc.RA119.010857. [Epub ahead of print]

The GATOR2-mTORC2 axis mediates Sestrin2-induced AKT Ser/Thr kinase activation.

Allison Ho Kowalsky, Sim Namkoong, Eric Mettetal, Hwan-Woo Park, Dubek Kazyken, Diane C Fingar, Jun Hee Lee.

  Sestrins represent a family of stress-inducible proteins that prevent the progression of many age- and obesity-associated disorders. Endogenous Sestrins maintain insulin-dependent AKT Ser/Thr kinase (AKT) activation during high-fat diet (HFD)-induced obesity, and overexpressed Sestrins activate AKT in various cell types, including liver and skeletal muscle cells. Although Sestrin-mediated AKT activation improves metabolic parameters, the mechanistic details underlying such improvement remain elusive. Here, we investigated how Sestrin2, the Sestrin homolog highly expressed in liver, induces strong AKT activation. We found that two known targets of Sestrin2, mTOR complex 1 (mTORC1) and AMP-activated protein kinase (AMPK), are not required for Sestrin2-induced AKT activation. Rather, phosphoinositol-3-kinase (PI3K) and mTORC2, kinases upstream of AKT, were essential for Sestrin2-induced AKT activation. Among these kinases, mTORC2 catalytic activity was strongly upregulated upon Sestrin2 overexpression in an in vitro kinase assay, indicating that mTORC2 may represent the major link between Sestrin2 and AKT. As reported previously, Sestrin2 interacted with mTORC2; however, we found here that this interaction occurs indirectly through GATOR2, a pentameric protein complex that directly interacts with Sestrin2. Deleting or silencing WD repeat domain 24 (WDR24), the GATOR2 component essential for the Sestrin2-GATOR2 interaction, or WDR59, the GATOR2 component essential for the GATOR2-mTORC2 interaction, completely ablated Sestrin2-induced AKT activation. We also noted that Sestrin2 also directly binds to the pleckstrin homology (PH) domain of AKT and induces AKT translocation to the plasma membrane. These results uncover a signaling mechanism whereby Sestrin2 activates AKT through GATOR2 and mTORC2.

Keywords:  Akt PKB; insulin resistance; liver metabolism; mTOR complex (mTORC); signal transduction

DOI:  https://doi.org/10.1074/jbc.RA119.010857
Front Cell Dev Biol. 2019 ;7 337

mTORC1-Rps15 Axis Contributes to the Mechanisms Underlying Global Translation Reduction During Senescence of Mouse Embryonic Fibroblasts.

Su Wu, Siyao Xu, Ruofei Li, Kecheng Li, Xiaoqin Zhong, Yingying Li, Zhifen Zhou, Yi Liu, Ran Feng, Jianfei Zheng, Zhou Songyang, Feng Liu.

  The reduction of protein translation is a common feature in senescent cells and aging organisms, yet the underlying mechanisms are not fully understood. Here we show that both global mRNA translation and mammalian/mechanistic target of rapamycin complex 1 (mTORC1) kinase activity are declined in a senescent model of mouse embryonic fibroblasts (MEFs). Furthermore, RNA-seq analyses from polysomal versus total mRNA fractions identify TOP-like mRNA of Rps15 whose translation is regulated by mTORC1 during MEF senescence. Overexpression of Rps15 delays MEF senescence, possibly through regulating ribosome maturation. Together, these findings indicate that the activation of mTORC1-Rps15 axis ameliorate senescence by regulating ribosome biogenesis, which may provide further insights into aging research.

Keywords:  MEF; Rps15; cell senescence; mRNA translation; mTORC1

DOI:  https://doi.org/10.3389/fcell.2019.00337
Curr Opin Cell Biol. 2019 Dec 31. pii: S0955-0674(19)30111-5. [Epub ahead of print]63 1-10

The ATG conjugation systems in autophagy.

Noboru Mizushima.

  Autophagosome formation and maturation involve the two ubiquitin-like systems: The ATG8 and ATG12 systems. ATG8 (LC3s and gamma-aminobutyric acid receptor-associated proteins in mammals) and ATG12 are covalently conjugated to phosphatidylethanolamine and ATG5, respectively. Although the ATG12 and ATG8 systems were discovered more than 20 years ago, their molecular functions are not fully understood. The aim of this review is to summarize recent findings related to ATG conjugation systems, focusing on current controversies regarding the genetic hierarchy of these systems, interpretation of conjugation-independent alternative macroautophagy, the differences in roles between LC3s and gamma-aminobutyric acid receptor-associated proteins in autophagosome formation and cargo recognition, and evolution of these systems.

Keywords:  ATG12; ATG8; Autophagosome; Ubiquitin

DOI:  https://doi.org/10.1016/j.ceb.2019.12.001
IEEE/ACM Trans Comput Biol Bioinform. 2020 Jan 06.

Exploring Molecular Descriptors and Fingerprints to Predict mTOR Kinase Inhibitors using Machine Learning Techniques.

Chetna Kumari, Muhammad Abulaish, Naidu Subbarao.

Mammalian Target of Rapamycin (mTOR) is a Ser/Thr protein kinase, and its role is integral to the autophagy pathway in cancer. Targeting mTOR for therapeutic interventions in cancer through autophagy pathway is challenging due to the dual roles of autophagy in tumor progression. The architecture of mTOR reveals two complexes - mTORC1 and mTORC2, each having multiple protein subunits. mTOR kinase inhibitors target the structurally and functionally similar catalytic subunits of both mTORC1 and mTORC2. In this paper, we have explored two different categories of molecular features - descriptors and fingerprints for developing predictive models using machine learning techniques. Random Forest variable importance measures and autoencoders are used to identify molecular descriptors and fingerprints, respectively. We have built various predictive models using identified features and their combination for predicting mTOR kinase inhibitors. Finally, the best model based on the Mathew correlation co-efficient value over the validation dataset is selected for screening kinase SARfari bioactivity dataset. In this study, we have identified twenty best performing descriptors for predicting mTOR kinase inhibitors. To the best of our knowledge, it is the first study on integrating traditional machine learning and deep learning-based approaches for feature extraction to predict mTOR kinase inhibitors.

DOI: https://doi.org/10.1109/TCBB.2020.2964203
Elife. 2020 Jan 08. pii: e50843. [Epub ahead of print]9

Cell-type specific regulation of neuronal intrinsic excitability by macroautophagy.

Ori J Lieberman, Micah D Frier, Avery F McGuirt, Christopher J Griffey, Elizabeth Rafikian, Mu Yang, Ai Yamamoto, Anders Borgkvist, Emanuela Santini, David Sulzer.

  The basal ganglia are a group of subcortical nuclei that contribute to action selection and reinforcement learning. The principal neurons of the striatum, spiny projection neurons of the direct (dSPN) and indirect (iSPN) pathways, maintain low intrinsic excitability, requiring convergent excitatory inputs to fire. Here, we examined the role of autophagy in mouse SPN physiology and animal behavior by generating conditional knockouts of Atg7 in either dSPNs or iSPNs. Loss of autophagy in either SPN population led to changes in motor learning but distinct effects on cellular physiology. dSPNs, but not iSPNs, required autophagy for normal dendritic structure and synaptic input. In contrast, iSPNs, but not dSPNs, were intrinsically hyperexcitable due to reduced function of the inwardly rectifying potassium channel, Kir2. These findings define a novel mechanism by which autophagy regulates neuronal activity: control of intrinsic excitability via the regulation of potassium channel function.

Keywords:  cell biology; mouse; neuroscience

DOI:  https://doi.org/10.7554/eLife.50843
Autophagy. 2020 Jan 07. 1-2

UVRAG in autophagy, inflammation, and cancer.

Ying Song, Christine Quach, Chengyu Liang.

  Macroautophagy/autophagy deregulation has been observed in perpetuated inflammation and the proliferation of tumor cells. However, the mechanisms underlying these changes have yet to be well-identified. UVRAG is one of the key players of autophagy, but its role in vivo remained puzzling. Our recent study utilized a mouse model with inducible expression of a cancer-derived frameshift (FS) mutation in UVRAG that dominant-negatively inhibits wild-type UVRAG, resulting in impaired stimulus-induced autophagy. The systemically compromised autophagy, particularly mitophagy, notably increases inflammation and associated pathologies. Furthermore, our discovery indicates that time-dependent autophagy suppression and ensuing CTNNB1/β-catenin activation may serve as one tumor-promoting mechanism underpinning age-related cancer susceptibility.

Keywords:  Autophagy; UVRAG; centrosome; inflammation; tumorigenesis; β-catenin

DOI:  https://doi.org/10.1080/15548627.2019.1709768
FASEB J. 2020 Jan;34(1): 161-179

Fas-apoptotic inhibitory molecule 2 localizes to the lysosome and facilitates autophagosome-lysosome fusion through the LC3 interaction region motif-dependent interaction with LC3.

Caroline Jeeyeon Hong, Jihye Yeon, Bo Kyoung Yeo, Hanwoong Woo, Hyun-Kyu An, Woojung Heo, Kyuhyung Kim, Seong-Woon Yu.

  Fas-apoptotic inhibitory molecule 2 (FAIM2) is a member of the transmembrane BAX inhibitor motif-containing (TMBIM) family. TMBIM family is comprised of six anti-apoptotic proteins that suppress cell death by regulating endoplasmic reticulum Ca2+ homeostasis. Recent studies have implicated two TMBIM proteins, GRINA and BAX Inhibitor-1, in mediating cytoprotection via autophagy. However, whether FAIM2 plays a role in autophagy has been unknown. Here we show that FAIM2 localizes to the lysosomes at basal state and facilitates autophagy through interaction with microtubule-associated protein 1 light chain 3 proteins in human neuroblastoma SH-SY5Y cells. FAIM2 overexpression increased autophagy flux, while autophagy flux was impaired in shRNA-mediated knockdown (shFAIM2) cells, and the impairment was more evident in the presence of rapamycin. In shFAIM2 cells, autophagosome maturation through fusion with lysosomes was impaired, leading to accumulation of autophagosomes. A functional LC3-interacting region motif within FAIM2 was essential for the interaction with LC3 and rescue of autophagy flux in shFAIM2 cells while LC3-binding property of FAIM2 was dispensable for the anti-apoptotic function in response to Fas receptor-mediated apoptosis. Suppression of autophagosome maturation was also observed in a null mutant of Caenorhabditis elegans lacking xbx-6, the ortholog of FAIM2. Our study suggests that FAIM2 is a novel regulator of autophagy mediating autophagosome maturation through the interaction with LC3.

Keywords:   xbx‐6 ; FAIM2; LIR; MAP1LC3B; autolysosome; autophagosome maturation

DOI:  https://doi.org/10.1096/fj.201901626R
Proc Natl Acad Sci U S A. 2020 Jan 09. pii: 201902788. [Epub ahead of print]

Innate immune receptor NOD2 mediates LGR5+ intestinal stem cell protection against ROS cytotoxicity via mitophagy stimulation.

Antonin Levy, Aline Stedman, Eric Deutsch, Françoise Donnadieu, Herbert W Virgin, Philippe J Sansonetti, Giulia Nigro.

  The nucleotide-binding oligomerization domain-containing protein 2 (NOD2) agonist muramyl dipeptide (MDP), a peptidoglycan motif common to all bacteria, supports leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5)+ intestinal stem cell (ISC) survival through NOD2 activation upon an otherwise lethal oxidative stress-mediated signal. However, the underlying protective mechanisms remain unknown. Here, using irradiation as stressor and primarily murine-derived intestinal organoids as a model system, we show that MDP induced a significant reduction of total and mitochondrial reactive oxygen species (ROS) within ISCs, which was associated with mitophagy induction. ATG16L1 knockout (KO) and NOD2 KO organoids did not benefit from the MDP-induced cytoprotection. We confirmed the MDP-dependent induction of ISC mitophagy upon stress in vivo. These findings elucidate the NOD2-mediated mechanism of cytoprotection involving the clearance of the lethal excess of ROS molecules through mitophagy, triggered by the coordinated activation of NOD2 and ATG16L1 by a nuclear factor κB (NF-κB)-independent pathway.

Keywords:  LGR5 intestinal stem cells; NOD2; ROS; autophagy; muramyl dipeptide

DOI:  https://doi.org/10.1073/pnas.1902788117
Mech Ageing Dev. 2020 Jan 07. pii: S0047-6374(20)30001-4. [Epub ahead of print] 111207

Mitophagy and DNA Damage Signaling in Human Aging.

Mansi Babbar, Sambuddha Basu, Beimeng Yang, Deborah L Croteau, Vilhelm A Bohr.

  Aging is associated with multiple human pathologies. In the past few years mitochondrial homeostasis has been well corelated with age-related disorders and longevity. Mitochondrial homeostasis involves generation, biogenesis and removal of dysfunctional mitochondria via mitophagy. Mitophagy is regulated by various mitochondrial and extra-mitochondrial factors including morphology, oxidative stress and DNA damage. For decades, DNA damage and inefficient DNA repair have been considered major determinants for age-related disorders. Although defects in DNA damage recognition and repair and mitophagy are well documented to be major factors in age-associated diseases, interactivity between these is poorly understood. Mitophagy efficiency decreases with age leading to accumulation of dysfunctional mitochondria enhancing the severity of age-related disorders including neurodegenerative diseases, inflammatory diseases, cancer, diabetes and many more. Therefore, mitophagy is being targeted for intervention in age-associated disorders. NAD+ supplementation has emerged as one intervention to target both defective DNA repair and mitophagy. In this review, we discuss the molecular signaling pathways involved in regulation of DNA damage and repair and of mitophagy, and we highlight the opportunities for clinical interventions targeting these processes to improve the quality of life during aging.

Keywords:  DNA damage: DNA Repair; aging; mitochondria; mitophagy

DOI:  https://doi.org/10.1016/j.mad.2020.111207
Commun Biol. 2019 Jan 25. 2(1): 37

Beclin 1 regulates recycling endosome and is required for skin development in mice.

Saori Noguchi, Shinya Honda, Tatsuya Saitoh, Hiroyuki Matsumura, Emi Nishimura, Shizuo Akira, Shigeomi Shimizu.

Beclin 1 is a key regulator of autophagy and endocytosis. However, its autophagy-independent functions remain poorly understood. Here, we report that Beclin 1 regulates recycling endosome and is required for skin development in vivo. We first established keratinocyte-specific Beclin 1-knockout mice and found that these mutant mice died owing to severe impairment of epidermal barrier. Beclin 1 plays a role in autophagy and the endocytic pathway in cooperation with Atg14 and UVRAG, respectively, and keratinocyte-specific Atg14-knockout mice do not show any abnormal phenotypes, suggesting that Beclin 1 has a role in skin development via the endocytic pathway. Furthermore, we found that Beclin 1 deficiency causes mislocalization of integrins via a defect of recycling endosome, abnormal cell detachment of basal cells and their immature differentiation, and abnormal skin development. These results provide the first genetic evidence showing the roles of Beclin 1 in recycling endosome and skin development.

DOI: https://doi.org/10.1038/s42003-018-0279-0
Adv Sci (Weinh). 2020 Jan;7(1): 1901261

TRIM14 Promotes Noncanonical NF-κB Activation by Modulating p100/p52 Stability via Selective Autophagy.

Meixin Chen, Zhiyao Zhao, Qingcai Meng, Puping Liang, Zexiong Su, Yaoxing Wu, Junjiu Huang, Jun Cui.

  The noncanonical NF-κB signaling pathway plays a critical role in a variety of biological functions including chronic inflammation and tumorigenesis. Activation of noncanonical NF-κB signaling largely relies on the abundance as well as the processing of the NF-κB family member p100/p52. Here, TRIM14 is identified as a novel positive regulator of the noncanonical NF-κB signaling pathway. TRIM14 promotes noncanonical NF-κB activation by targeting p100/p52 in vitro and in vivo. Furthermore, a mechanistic study shows that TRIM14 recruits deubiquitinase USP14 to cleave the K63-linked ubiquitin chains of p100/p52 at multiple sites, thereby preventing p100/p52 from cargo receptor p62-mediated autophagic degradation. TRIM14 deficiency in mice significantly impairs noncanonical NF-κB-mediated inflammatory responses as well as acute colitis and colitis-associated colon cancer development. Taken together, these findings establish the TRIM14-USP14 axis as a crucial checkpoint that controls noncanonical NF-κB signaling and highlight the crosstalk between autophagy and innate immunity.

Keywords:  TRIM14; inflammation; noncanonical NF‐κB signaling; p100/p52; selective autophagy

DOI:  https://doi.org/10.1002/advs.201901261
FASEB J. 2019 Dec 13.

p53 regulates mitochondrial dynamics by inhibiting Drp1 translocation into mitochondria during cellular senescence.

Young Yeon Kim, Jee-Hyun Um, Jeong-Hyun Yoon, Da-Ye Lee, Yoon Jung Lee, Dong Hyun Kim, Joo-In Park, Jeanho Yun.

  Cellular senescence acts as an important barrier to tumorigenesis by eliminating precancerous cells. Previous studies have shown an essential role of the tumor suppressor p53 in cellular senescence, but how p53 induces cellular senescence is not fully understood. We found that p53 promoted the formation of highly interconnected and elongated mitochondria prior to the onset of cellular senescence. The inhibition of mitochondrial elongation upon p53 expression suppressed cellular senescence, suggesting that mitochondrial elongation is required for the induction of p53-dependent senescence. p53-induced mitochondrial elongation resulted in mitochondrial dysfunction and subsequent increases in intracellular reactive oxygen species (ROS) levels, an important mediator of cellular senescence. Mechanistically, the inhibitory phosphorylation of Drp1 Ser637 increased upon p53 expression, suppressing the translocation of Drp1 into mitochondria. The transcriptional function of p53 was crucial for controlling the inhibitory phosphorylation of Drp1, whereas p21 was nonessential. Protein kinase A (PKA) activity was responsible for p53-mediated Drp1 Ser637 phosphorylation and mitochondrial dysfunction. Taken together, these results suggest that p53 regulates mitochondrial dynamics through the PKA-Drp1 pathway to induce cellular senescence.

Keywords:  Drp1; PKA; cellular senescence; mitochondria dynamics; p53; senescence

DOI:  https://doi.org/10.1096/fj.201901747RR
Stem Cell Reports. 2019 Dec 20. pii: S2213-6711(19)30444-8. [Epub ahead of print]

Pathogenic Pathways in Early-Onset Autosomal Recessive Parkinson's Disease Discovered Using Isogenic Human Dopaminergic Neurons.

Tim Ahfeldt, Alban Ordureau, Christina Bell, Lily Sarrafha, Chicheng Sun, Silvia Piccinotti, Tobias Grass, Gustavo M Parfitt, Joao A Paulo, Fumiki Yanagawa, Takayuki Uozumi, Yasujiro Kiyota, J Wade Harper, Lee L Rubin.

  Parkinson's disease (PD) is a complex and highly variable neurodegenerative disease. Familial PD is caused by mutations in several genes with diverse and mostly unknown functions. It is unclear how dysregulation of these genes results in the relatively selective death of nigral dopaminergic neurons (DNs). To address this question, we modeled PD by knocking out the PD genes PARKIN (PRKN), DJ-1 (PARK7), and ATP13A2 (PARK9) in independent isogenic human pluripotent stem cell (hPSC) lines. We found increased levels of oxidative stress in all PD lines. Increased death of DNs upon differentiation was found only in the PARKIN knockout line. Using quantitative proteomics, we observed dysregulation of mitochondrial and lysosomal function in all of the lines, as well as common and distinct molecular defects caused by the different PD genes. Our results suggest that precise delineation of PD subtypes will require evaluation of molecular and clinical data.

Keywords:  ATP13A2; CRISPR; DJ1; Parkin; Parkinson's disease; disease modeling; genome editing; human pluripotent stem cells; proteomics; transcriptomics

DOI:  https://doi.org/10.1016/j.stemcr.2019.12.005
Exp Neurobiol. 2019 Dec 31. 28(6): 643-657

Autophagy, Cellular Aging and Age-related Human Diseases.

So Yeong Cheon, Hyunjeong Kim, David C Rubinsztein, Jong Eun Lee.

  Macroautophagy/autophagy is a conserved degradation system that engulfs intracytoplasmic contents, including aggregated proteins and organelles, which is crucial for cellular homeostasis. During aging, cellular factors suggested as the cause of aging have been reported to be associated with progressively compromised autophagy. Dysfunctional autophagy may contribute to age-related diseases, such as neurodegenerative disease, cancer, and metabolic syndrome, in the elderly. Therefore, restoration of impaired autophagy to normal may help to prevent age-related disease and extend lifespan and longevity. Therefore, this review aims to provide an overview of the mechanisms of autophagy underlying cellular aging and the consequent disease. Understanding the mechanisms of autophagy may provide potential information to aid therapeutic interventions in age-related diseases.

Keywords:  Aging; Autophagy; DNA damage; Oxidative stress; SASP; Telomere shortening

DOI:  https://doi.org/10.5607/en.2019.28.6.643
Mol Neurodegener. 2020 Jan 06. 15(1): 2

Promoting tau secretion and propagation by hyperactive p300/CBP via autophagy-lysosomal pathway in tauopathy.

Xu Chen, Yaqiao Li, Chao Wang, Yinyan Tang, Sue-Ann Mok, Richard M Tsai, Julio C Rojas, Anna Karydas, Bruce L Miller, Adam L Boxer, Jason E Gestwicki, Michelle Arkin, Ana Maria Cuervo, Li Gan.

   BACKGROUND: The trans-neuronal propagation of tau has been implicated in the progression of tau-mediated neurodegeneration. There is critical knowledge gap in understanding how tau is released and transmitted, and how that is dysregulated in diseases. Previously, we reported that lysine acetyltransferase p300/CBP acetylates tau and regulates its degradation and toxicity. However, whether p300/CBP is involved in regulation of tau secretion and propagation is unknown.
METHOD: We investigated the relationship between p300/CBP activity, the autophagy-lysosomal pathway (ALP) and tau secretion in mouse models of tauopathy and in cultured rodent and human neurons. Through a high-through-put compound screen, we identified a new p300 inhibitor that promotes autophagic flux and reduces tau secretion. Using fibril-induced tau spreading models in vitro and in vivo, we examined how p300/CBP regulates tau propagation.
RESULTS: Increased p300/CBP activity was associated with aberrant accumulation of ALP markers in a tau transgenic mouse model. p300/CBP hyperactivation blocked autophagic flux and increased tau secretion in neurons. Conversely, inhibiting p300/CBP promoted autophagic flux, reduced tau secretion, and reduced tau propagation in fibril-induced tau spreading models in vitro and in vivo.
CONCLUSIONS: We report that p300/CBP, a lysine acetyltransferase aberrantly activated in tauopathies, causes impairment in ALP, leading to excess tau secretion. This effect, together with increased intracellular tau accumulation, contributes to enhanced spreading of tau. Our findings suggest that inhibition of p300/CBP as a novel approach to correct ALP dysfunction and block disease progression in tauopathy.

Keywords:  Autophagy-lysosomal pathway; Tau secretion; Tau spreading; Tauopathy; p300/CBP

DOI:  https://doi.org/10.1186/s13024-019-0354-0
Front Oncol. 2019 ;9 1373

mTORC1 as a Regulator of Mitochondrial Functions and a Therapeutic Target in Cancer.

Karen Griselda de la Cruz López, Mariel Esperanza Toledo Guzmán, Elizabeth Ortiz Sánchez, Alejandro García Carrancá.

  Continuous proliferation of tumor cells requires constant adaptations of energy metabolism to rapidly fuel cell growth and division. This energetic adaptation often comprises deregulated glucose uptake and lactate production in the presence of oxygen, a process known as the "Warburg effect." For many years it was thought that the Warburg effect was a result of mitochondrial damage, however, unlike this proposal tumor cell mitochondria maintain their functionality, and is essential for integrating a variety of signals and adapting the metabolic activity of the tumor cell. The mammalian/mechanistic target of rapamycin complex 1 (mTORC1) is a master regulator of numerous cellular processes implicated in proliferation, metabolism, and cell growth. mTORC1 controls cellular metabolism mainly by regulating the translation and transcription of metabolic genes, such as peroxisome proliferator activated receptor γ coactivator-1 α (PGC-1α), sterol regulatory element-binding protein 1/2 (SREBP1/2), and hypoxia inducible factor-1 α (HIF-1α). Interestingly it has been shown that mTORC1 regulates mitochondrial metabolism, thus representing an important regulator in mitochondrial function. Here we present an overview on the role of mTORC1 in the regulation of mitochondrial functions in cancer, considering new evidences showing that mTORC1 regulates the translation of nucleus-encoded mitochondrial mRNAs that result in an increased ATP mitochondrial production. Moreover, we discuss the relationship between mTORC1 and glutaminolysis, as well as mitochondrial metabolites. In addition, mitochondrial fission processes regulated by mTORC1 and its impact on cancer are discussed. Finally, we also review the therapeutic efficacy of mTORC1 inhibitors in cancer treatments, considering its use in combination with other drugs, with particular focus on cellular metabolism inhibitors, that could help improve their anti neoplastic effect and eliminate cancer cells in patients.

Keywords:  cancer; mTORC1; mitochondria; mitochondrial functions; therapy

DOI:  https://doi.org/10.3389/fonc.2019.01373
Autophagy. 2020 Jan 10. 1-25

SQSTM1/p62 activates NFE2L2/NRF2 via ULK1-mediated autophagic KEAP1 degradation and protects mouse liver from lipotoxicity.

Da Hyun Lee, Jeong Su Park, Yu Seol Lee, Jisu Han, Dong-Kyu Lee, Sung Won Kwon, Dai Hoon Han, Yong-Ho Lee, Soo Han Bae.

  Lipotoxicity, induced by saturated fatty acid (SFA)-mediated cell death, plays an important role in the pathogenesis of nonalcoholic fatty liver disease (NAFLD). The KEAP1 (kelch like ECH associated protein 1)-NFE2L2/NRF2 (nuclear factor, erythroid 2 like 2) pathway is a pivotal defense mechanism against lipotoxicity. We previously reported that SQSTM1/p62 has a cytoprotective role against lipotoxicity through activation of the noncanonical KEAP1- NFE2L2 pathway in hepatocytes. However, the underlying mechanisms and physiological relevance of this pathway have not been clearly defined. Here, we demonstrate that NFE2L2-mediated induction of SQSTM1 activates the noncanonical KEAP1-NFE2L2 pathway under lipotoxic conditions. Furthermore, we identified that SQSTM1 induces ULK1 (unc-51 like autophagy activating kinase 1) phosphorylation by facilitating the interaction between AMPK (AMP-activated protein kinase) and ULK1, leading to macroautophagy/autophagy induction, followed by KEAP1 degradation and NFE2L2 activation. Accordingly, the activity of this SQSTM1-mediated noncanonical KEAP1-NFE2L2 pathway conferred hepatoprotection against lipotoxicity in the livers of conventional sqstm1- and liver-specific sqstm1-knockout mice. Moreover, this pathway activity was evident in the livers of patients with nonalcoholic fatty liver. This axis could thus represent a novel target for NAFLD treatment.Abbreviations: ACACA: acetyl-CoA carboxylase alpha; ACTB: actin beta; BafA1: bafilomycin A1; CM-H2DCFDA:5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate; CQ: chloroquine; CUL3: cullin 3; DMSO: dimethyl sulfoxide; FASN: fatty acid synthase; GSTA1: glutathione S-transferase A1; HA: hemagglutinin; Hepa1c1c7: mouse hepatoma cells; HMOX1/HO-1: heme oxygenase 1; KEAP1: kelch like ECH associated protein 1; MAP1LC3B/LC3B: microtubule-associated protein 1 light chain 3; MEF: mouse embryonic fibroblast; MTORC1: mechanistic target of rapamycin kinase complex 1; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; NAC: N-acetyl-L-cysteine; NAFLD: nonalcoholic fatty liver disease; NASH: nonalcoholic steatohepatitis; NFE2L2/NRF2: nuclear factor, erythroid 2 like 2; NQO1: NAD(P)H quinone dehydrogenase 1; PA: palmitic acid; PARP: poly (ADP-ribose) polymerase 1; PRKAA1/2: protein kinase AMP-activated catalytic subunits alpha1/2; RBX1: ring-box 1; ROS: reactive oxygen species; SESN2: sestrin 2; SFA: saturated fatty acid; siRNA: small interfering RNA; SQSTM1/p62: sequestosome 1; SREBF1: sterol regulatory element binding transcription factor 1; TBK1: TANK binding kinase 1; TUNEL: terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling; ULK1: unc-51 like autophagy activating kinase.

Keywords:  KEAP1-NFE2L2 pathway; NAFLD; SQSTM1; ULK1; lipotoxicity

DOI:  https://doi.org/10.1080/15548627.2020.1712108
J Biol Chem. 2020 Jan 09. pii: jbc.RA119.010912. [Epub ahead of print]

Use of the LC3B-fusion technique for biochemical and structural studies of proteins involved in the N-degron pathway.

Leehyeon Kim, Do Hoon Kwon, Jiwon Heo, Mi Rae Park, Hyun Kyu Song.

  The N-degron pathway, formerly the N-end rule pathway, is a protein degradation process that determines the half-life of proteins based on their N-terminal residues. In contrast to the well-established in vivo studies over decades, in vitro studies of this pathway, including biochemical characterization and high-resolution structures, are relatively limited. In this study, we have developed a unique fusion technique using microtubule-associated protein 1A/1B light chain 3B, a key marker protein of autophagy, to tag the N-terminus of the proteins involved in the N-degron pathway, which enables high yield of homogeneous target proteins with variable N-terminal residues for diverse biochemical studies including enzymatic and binding assays and substrate identification. Intriguingly, crystallization showed a markedly enhanced probability, even for the N-degron complexes. To validate our results, we determined the structures of select proteins in the N-degron pathway and compared them to the PDB-deposited proteins. Furthermore, several biochemical applications of this technique were introduced. Therefore, this technique can be used as a general tool for the in vitro study of the N-degron pathway.

Keywords:  LC3B; N-end rule; NTAQ1; PRT1; UBR box; autophagy; p62 (sequestosome 1(SQSTM1)); protein crystallization; protein degradation; ubiquitylation (ubiquitination)

DOI:  https://doi.org/10.1074/jbc.RA119.010912
Trends Neurosci. 2020 Jan 06. pii: S0166-2236(19)30224-3. [Epub ahead of print]

The Sigma-1 Receptor at the Crossroad of Proteostasis, Neurodegeneration, and Autophagy.

Maximilian G Christ, Albrecht M Clement, Christian Behl.

  Neurodegenerative diseases are linked to dysfunctional proteostasis and disturbed autophagy. Here, we discuss how the sigma-1 receptor (Sig-1R) may act at the intersection of this interaction, as loss-of-function mutations of this unique chaperone are associated with defective autophagy and its pharmacological activation induces autophagic activity.

Keywords:  ALS; ER; autophagy; neurodegeneration; sigma-1 receptor

DOI:  https://doi.org/10.1016/j.tins.2019.12.002
J Biol Chem. 2020 Jan 07. pii: jbc.RA119.009699. [Epub ahead of print]

A dual druggable genome-wide siRNA and compound library screening approach identifies modulators of parkin recruitment to mitochondria.

Helen L Scott, Nicola Buckner, Francesc Fernandez-Albert, Elisa Pedone, Lorena Postiglione, Gongyu Shi, Nicholas Allen, Liang-Fong Wong, Lorenzo Magini, Lucia Marucci, Gregory O'Sullivan, Sarah Cole, Justin Powell, Peter Maycox, James B Uney.

  Genetic and biochemical evidence points to an association between mitochondrial dysfunction and Parkinson's disease (PD). PD-associated mutations in several genes have been identified and include those encoding PTEN-induced putative kinase 1 (PINK1) and parkin. To identify genes, pathways, and pharmacological targets that modulate the clearance of damaged or old mitochondria (mitophagy), here we developed a high-content imaging-based assay of parkin recruitment to mitochondria and screened both a druggable genome-wide siRNA library and a small neuroactive compound library. We used a multi-parameter principal component analysis and an unbiased parameter-agnostic machine-learning approach to analyze the siRNA-based screening data. The hits identified in this analysis included specific genes of the ubiquitin proteasome system, and inhibition of ubiquitin-conjugating enzyme 2 N (UBE2N) with a specific antagonist, Bay 11-7082, indicated that UBE2N modulates parkin recruitment and downstream events in the mitophagy pathway. Screening of the compound library identified kenpaullone, an inhibitor of cyclin-dependent kinases (CDKs) and glycogen synthase kinase 3 (GSK3), as a modulator of parkin recruitment. Validation studies revealed that kenpaullone augments the mitochondrial network and protects against the complex I inhibitor MPP+. Finally, we used a microfluidics platform to assess the timing of parkin recruitment to depolarized mitochondria and its modulation by kenpaullone in real time and with single-cell resolution. We demonstrate that the high-content imaging-based assay presented here is suitable for both genetic and pharmacological screening approaches, and we also provide evidence that pharmacological compounds modulate PINK1-dependent parkin recruitment.

Keywords:  Kenpaullone; Parkinson disease; mitochondria; mitophagy; parkin; ubiquitin

DOI:  https://doi.org/10.1074/jbc.RA119.009699
Autophagy. 2020 Jan 10.

Autophagy-Dependent Ferroptosis Drives Tumor-Associated Macrophage Polarization via Release and Uptake of Oncogenic KRAS Protein.

Enyong Dai, Leng Han, Jiao Liu, Yangchun Xie, Guido Kroemer, Daniel J Klionsky, Herbert J Zeh, Rui Kang, Jing Wang, Daolin Tang.

  KRAS is the most frequently mutated oncogene in human neoplasia. Despite a large investment to understand the effects of KRAS mutation in cancer cells, the direct effects of the oncogenetic KRAS activation on immune cells remain elusive. Here, we report that extracellular KRASG12D is essential for pancreatic tumor-associated macrophage polarization. Oxidative stress induces KRASG12D protein release from cancer cells succumbing to autophagy-dependent ferroptosis. Extracellular KRASG12D packaged into exosomes then is taken up by macrophages through an AGER-dependent mechanism. KRASG12D causes macrophages to switch to an M2-like pro-tumor phenotype via STAT3-dependent fatty acid oxidation. Consequently, the disruption of KRASG12D release and uptake can abolish the macrophage-mediated stimulation of pancreatic adenocarcinomas in mouse models. Importantly, the level of KRASG12D expression in macrophages correlates with poor survival in pancreatic cancer patients. These findings not only identify extracellular KRASG12D as a key mediator of cancer cell-macrophage communication, but also provide a novel KRAS-targeted anticancer strategy.

Keywords:  AGER; DAMP; KRAS; autophagy; exosomes; ferroptosis; macrophage; pancreatic cancer

DOI:  https://doi.org/10.1080/15548627.2020.1714209
Commun Biol. 2019 Sep 18. 2(1): 346

Inhibition of the glutamine transporter SNAT1 confers neuroprotection in mice by modulating the mTOR-autophagy system.

Daisuke Yamada, Kenji Kawabe, Ikue Tosa, Shunpei Tsukamoto, Ryota Nakazato, Miki Kou, Koichi Fujikawa, Saki Nakamura, Mitsuaki Ono, Toshitaka Oohashi, Mari Kaneko, Shioi Go, Eiichi Hinoi, Yukio Yoneda, Takeshi Takarada.

The pathophysiological role of mammalian target of rapamycin complex 1 (mTORC1) in neurodegenerative diseases is established, but possible therapeutic targets responsible for its activation in neurons must be explored. Here we identified solute carrier family 38a member 1 (SNAT1, Slc38a1) as a positive regulator of mTORC1 in neurons. Slc38a1flox/flox and Synapsin I-Cre mice were crossed to generate mutant mice in which Slc38a1 was selectively deleted in neurons. Measurement of 2,3,5-triphenyltetrazolium chloride (TTC) or the MAP2-negative area in a mouse model of middle cerebral artery occlusion (MCAO) revealed that Slc38a1 deficiency decreased infarct size. We found a transient increase in the phosphorylation of p70S6k1 (pp70S6k1) and a suppressive effect of rapamycin on infarct size in MCAO mice. Autophagy inhibitors completely mitigated the suppressive effect of SNAT1 deficiency on neuronal cell death under in vitro stroke culture conditions. These results demonstrate that SNAT1 promoted ischemic brain damage via mTOR-autophagy system.

DOI: https://doi.org/10.1038/s42003-019-0582-4
Thorac Cancer. 2020 Jan 08.

Transcriptional regulation of autophagy-lysosomal pathway in cancer.

Xinzhong Chang, Ruihua Dong.

  The transcriptional regulation of autophagy-lysosomal pathway adapts to cellular stress and enables advanced cancer cells survive. This pathway plays an oncopromoting or oncosuppressing role, depending on context-dependent stresses and treatment resistance. It remains controversial whether this pathway represents a target for drugs, although autophagy-lysosomal inducers and inhibitors have been tested in clinical trials for cancer treatment. Therefore, identifying the transcriptional regulators of autophagy-lysosomal pathway may lead to the development of effective cancer treatment and the improvement of the existing targeted cancer therapies. In this review, we summarize findings from several published studies on transcriptional regulation of autophagy-lysosomal pathway in cancer biology, and evaluate its functional role as a therapeutic target.

Keywords:  Autophagy; lysosome; oncoprotein; oncosuppressor; transcriptional regulator

DOI:  https://doi.org/10.1111/1759-7714.13287
Front Neurosci. 2019 ;13 1352

Autophagy and LRRK2 in the Aging Brain.

Federica Albanese, Salvatore Novello, Michele Morari.

  Autophagy is a highly conserved process by which long-lived macromolecules, protein aggregates and dysfunctional/damaged organelles are delivered to lysosomes for degradation. Autophagy plays a crucial role in regulating protein quality control and cell homeostasis in response to energetic needs and environmental challenges. Indeed, activation of autophagy increases the life-span of living organisms, and impairment of autophagy is associated with several human disorders, among which neurodegenerative disorders of aging, such as Parkinson's disease. These disorders are characterized by the accumulation of aggregates of aberrant or misfolded proteins that are toxic for neurons. Since aging is associated with impaired autophagy, autophagy inducers have been viewed as a strategy to counteract the age-related physiological decline in brain functions and emergence of neurodegenerative disorders. Parkinson's disease is a hypokinetic, multisystemic disorder characterized by age-related, progressive degeneration of central and peripheral neuronal populations, associated with intraneuronal accumulation of proteinaceous aggregates mainly composed by the presynaptic protein α-synuclein. α-synuclein is a substrate of macroautophagy and chaperone-mediated autophagy (two major forms of autophagy), thus impairment of its clearance might favor the process of α-synuclein seeding and spreading that trigger and sustain the progression of this disorder. Genetic factors causing Parkinson's disease have been identified, among which mutations in the LRRK2 gene, which encodes for a multidomain protein encompassing central GTPase and kinase domains, surrounded by protein-protein interaction domains. Six LRRK2 mutations have been pathogenically linked to Parkinson's disease, the most frequent being the G2019S in the kinase domain. LRRK2-associated Parkinson's disease is clinically and neuropathologically similar to idiopathic Parkinson's disease, also showing age-dependency and incomplete penetrance. Several mechanisms have been proposed through which LRRK2 mutations can lead to Parkinson's disease. The present article will focus on the evidence that LRRK2 and its mutants are associated with autophagy dysregulation. Studies in cell lines and neurons in vitro and in LRRK2 knock-out, knock-in, kinase-dead and transgenic animals in vivo will be reviewed. The role of aging in LRRK2-induced synucleinopathy will be discussed. Possible mechanisms underlying the LRRK2-mediated control over autophagy will be analyzed, and the contribution of autophagy dysregulation to the neurotoxic actions of LRRK2 will be examined.

Keywords:  LAMP2A; LC3; LRRK2; Parkinson’s disease; aging; autophagy; lysosomes; α-synuclein

DOI:  https://doi.org/10.3389/fnins.2019.01352
FASEB J. 2020 Jan;34(1): 399-409

Autophagy mediates hepatic GRK2 degradation to facilitate glucagon-induced metabolic adaptation to fasting.

Marta Cruces-Sande, Alba C Arcones, Rocío Vila-Bedmar, Almudena Val-Blasco, Kfir Sharabi, Daniel Díaz-Rodríguez, Pere Puigserver, Federico Mayor, Cristina Murga.

  The liver plays a key role during fasting to maintain energy homeostasis and euglycemia via metabolic processes mainly orchestrated by the insulin/glucagon ratio. We report here that fasting or calorie restriction protocols in C57BL6 mice promote a marked decrease in the hepatic protein levels of G protein-coupled receptor kinase 2 (GRK2), an important negative modulator of both G protein-coupled receptors (GPCRs) and insulin signaling. Such downregulation of GRK2 levels is liver-specific and can be rapidly reversed by refeeding. We find that autophagy, and not the proteasome, represents the main mechanism implicated in fasting-induced GRK2 degradation in the liver in vivo. Reducing GRK2 levels in murine primary hepatocytes facilitates glucagon-induced glucose production and enhances the expression of the key gluconeogenic enzyme Pck1. Conversely, preventing full downregulation of hepatic GRK2 during fasting using adenovirus-driven overexpression of this kinase in the liver leads to glycogen accumulation, decreased glycemia, and hampered glucagon-induced gluconeogenesis, thus preventing a proper and complete adaptation to nutrient deprivation. Overall, our data indicate that physiological fasting-induced downregulation of GRK2 in the liver is key for allowing complete glucagon-mediated responses and efficient metabolic adaptation to fasting in vivo.

Keywords:  GPCR; GRK2; autophagy; calorie restriction; fasting; glucagon signaling; gluconeogenesis; intermittent fasting

DOI:  https://doi.org/10.1096/fj.201901444R
Nat Commun. 2020 Jan 08. 11(1): 117

ULK1 phosphorylates Exo70 to suppress breast cancer metastasis.

Liyuan Mao, Yan-Yan Zhan, Bin Wu, Qiang Yu, Liang Xu, Xiaoting Hong, Linhai Zhong, Panying Mi, Li Xiao, Xinquan Wang, Hanwei Cao, Wenqing Zhang, Binbin Chen, Jingzhou Xiang, Kunrong Mei, Ravi Radhakrishnan, Wei Guo, Tianhui Hu.

Increased expression of protein kinase ULK1 was reported to negatively correlate with breast cancer metastasis. Here we report that ULK1 suppresses the migration and invasion of human breast cancer cells. The suppressive effect is mediated through direct phosphorylation of Exo70, a key component of the exocyst complex. ULK1 phosphorylation inhibits Exo70 homo-oligomerization as well as its assembly to the exocyst complex, which are needed for cell protrusion formation and matrix metalloproteinases secretion during cell invasion. Reversely, upon growth factor stimulation, Exo70 is phosphorylated by ERK1/2, which in turn suppresses its phosphorylation by ULK1. Together, our study identifies Exo70 as a substrate of ULK1 that inhibits cancer metastasis, and demonstrates that two counteractive regulatory mechanisms are well orchestrated during tumor cell invasion.

DOI: https://doi.org/10.1038/s41467-019-13923-7
Sci Adv. 2020 Jan;6(1): eaaz1441

UPRER promotes lipophagy independent of chaperones to extend life span.

Joseph R Daniele, Ryo Higuchi-Sanabria, Jenni Durieux, Samira Monshietehadi, Vidhya Ramachandran, Sarah U Tronnes, Naame Kelet, Melissa Sanchez, Melissa G Metcalf, Gilberto Garcia, Phillip A Frankino, Camila Benitez, Mandy Zeng, Daniel J Esping, Larry Joe, Andrew Dillin.

Longevity is dictated by a combination of environmental and genetic factors. One of the key mechanisms to regulate life-span extension is the induction of protein chaperones for protein homeostasis. Ectopic activation of the unfolded protein response of the endoplasmic reticulum (UPRER) specifically in neurons is sufficient to enhance organismal stress resistance and extend life span. Here, we find that this activation not only promotes chaperones but also facilitates ER restructuring and ER function. This restructuring is concomitant with lipid depletion through lipophagy. Activation of lipophagy is distinct from chaperone induction and is required for the life-span extension found in this paradigm. Last, we find that overexpression of the lipophagy component, ehbp-1, is sufficient to deplete lipids, remodel ER, and promote life span. Therefore, UPR induction in neurons triggers two distinct programs in the periphery: the proteostasis arm through protein chaperones and metabolic changes through lipid depletion mediated by EH domain binding protein 1 (EHBP-1).

DOI: https://doi.org/10.1126/sciadv.aaz1441
Proc Natl Acad Sci U S A. 2020 Jan 09. pii: 201918931. [Epub ahead of print]

Loss of TSC complex enhances gluconeogenesis via upregulation of Dlk1-Dio3 locus miRNAs.

Dritan Liko, Andrzej Rzepiela, Vanja Vukojevic, Mihaela Zavolan, Michael N Hall.

  Loss of the tumor suppressor tuberous sclerosis complex 1 (Tsc1) in the liver promotes gluconeogenesis and glucose intolerance. We asked whether this could be attributed to aberrant expression of small RNAs. We performed small-RNA sequencing on liver of Tsc1-knockout mice, and found that miRNAs of the delta-like homolog 1 (Dlk1)-deiodinase iodothyronine type III (Dio3) locus are up-regulated in an mTORC1-dependent manner. Sustained mTORC1 signaling during development prevented CpG methylation and silencing of the Dlk1-Dio3 locus, thereby increasing miRNA transcription. Deletion of miRNAs encoded by the Dlk1-Dio3 locus reduced gluconeogenesis, glucose intolerance, and fasting blood glucose levels. Thus, miRNAs contribute to the metabolic effects observed upon loss of TSC1 and hyperactivation of mTORC1 in the liver. Furthermore, we show that miRNA is a downstream effector of hyperactive mTORC1 signaling.

Keywords:  CpG methylation; Dlk1-Dio3; glucose metabolism; mTOR; miRNA

DOI:  https://doi.org/10.1073/pnas.1918931117