bims-polyam 2021-11-07 papers

bims-polyam

Biomed News

on Polyamines

Issue of 2021–11–07
seven papers selected by
Sebastian J. Hofer, University of Graz

Cryo-EM structures and transport mechanism of human P5B type ATPase ATP13A2.
Spermine protects aluminium chloride and iron-induced neurotoxicity in rat model of Alzheimer's disease via attenuation of tau phosphorylation, Amyloid-β (1-42) and NF-κB pathway.
Differential Association of Free, Conjugated, and Bound Forms of Polyamines and Transcript Abundance of Their Biosynthetic and Catabolic Genes During Drought/Salinity Stress in Tomato (Solanum lycopersicum L.) Leaves.
Structure of Pseudomonas aeruginosa spermidine dehydrogenase: a polyamine oxidase with a novel heme-binding fold.
Putrescine and its metabolic precursor arginine promote biofilm and c-di-GMP synthesis in Pseudomonas aeruginosa.
Exogenous Spermidine Priming Mitigates the Osmotic Damage in Germinating Seeds of Leymus chinensis Under Salt-Alkali Stress.
The endocytic pathway taken by cationic substances requires Rab14 but not Rab5 and Rab7.

Cell Discov. 2021 Nov 02. 7(1): 106

Cryo-EM structures and transport mechanism of human P5B type ATPase ATP13A2.

Xudong Chen, Mingze Zhou, Sensen Zhang, Jian Yin, Ping Zhang, Xujun Xuan, Peiyi Wang, Zhiqiang Liu, Boda Zhou, Maojun Yang.

Polyamines are important polycations that play critical roles in mammalian cells. ATP13A2 belongs to the orphan P5B adenosine triphosphatases (ATPase) family and has been established as a lysosomal polyamine exporter to maintain the normal function of lysosomes and mitochondria. Previous studies have reported that several human neurodegenerative disorders are related to mutations in the ATP13A2 gene. However, the transport mechanism of ATP13A2 in the lysosome remains unclear. Here, we report the cryo-electron microscopy (cryo-EM) structures of three distinct intermediates of the human ATP13A2, revealing key insights into the spermine (SPM) transport cycle in the lysosome. The transmembrane domain serves as a substrate binding site and the C-terminal domain is essential for protein stability and may play a regulatory role. These findings advance our understanding of the polyamine transport mechanism, the lipid-associated regulation, and the disease-associated mutants of ATP13A2.

DOI: https://doi.org/10.1038/s41421-021-00334-6
Inflammopharmacology. 2021 Nov 02.

Spermine protects aluminium chloride and iron-induced neurotoxicity in rat model of Alzheimer's disease via attenuation of tau phosphorylation, Amyloid-β (1-42) and NF-κB pathway.

Khadga Raj, G D Gupta, Shamsher Singh.

  Alzheimer's disease (AD) is the most prevalent type of dementia, characterized by a gradual decline in cognitive and memory functions of the aged peoples. Long-term exposure to heavy metals (aluminium and iron) cause neurotoxicity by amyloid plaques accumulation, tau phosphorylation, increased oxidative stress, neuroinflammation, and cholinergic neurons degeneration, contributes to the development of AD-like symptoms. The present research work is designed to investigate the neuroprotective effect of spermine in aluminium chloride (AlCl3), and iron (Fe) induced AD-like symptoms in rats. Rats were administered of AlCl3 (100 mg/kg p.o.) alone and in combination with iron (120 μg/g, p.o.) for 28 days. Spermine (5 and 10 mg/kg) through intraperitoneal (i.p.) route was given for 14 days. The recognition and spatial memory impairment were tasted using Morris water maze (MWM), actophotometer, and Novel Object Recognition test (NORT). All the rats were sacrificed on day 29, brains were isolated, and tissue homogenate was used for neuroinflammatory, biochemical, neurotransmitters, metals concentration, and nuclear factor-kappa B (NF-κB) analysis. In the present study, AlCl3 and iron administration elevated oxidative stress, cytokines release, dysbalanced neurotransmitters concentration, and biochemical changes. Rats treated with spermine dose-dependently improved the recognition and spatial memory, attenuated proinflammatory cytokine release, and restored neurotransmitters concentration and antioxidant enzymes. Spermine also mitigated the increased beta-amyloid (Aβ42), with downregulation of tau phosphorylation. Furthermore, spermine augmented the hippocampal levels of B cell leukaemia/lymphoma-2 (Bcl-2), diminished nuclear factor-kappa B (NF-κB) and caspase-3 (casp-3) expression. Moreover, spermine exhibited the neuroprotective effect through anti-inflammatory, antioxidant, neurotransmitters restoration, anti-apoptotic Aβ42 concentration.

Keywords:  Aluminium chloride; Alzheimer’s disease; Hippocampus; Iron; Neurotransmitters; Proinflammatory cytokines

DOI:  https://doi.org/10.1007/s10787-021-00883-y
Front Plant Sci. 2021 ;12 743568

Differential Association of Free, Conjugated, and Bound Forms of Polyamines and Transcript Abundance of Their Biosynthetic and Catabolic Genes During Drought/Salinity Stress in Tomato (Solanum lycopersicum L.) Leaves.

Rakesh K Upadhyay, Tahira Fatima, Avtar K Handa, Autar K Mattoo.

  Polyamines have been implicated in ameliorating the detrimental effects of drought and saline conditions on plant growth and development. The independent impact of these two abiotic stresses on polyamine (PA) biosynthesis, catabolism, and homeostasis, as well as on their transcript abundance in tomato leaves, is presented here. We show that the total levels of putrescine (PUT), spermidine (SPD), and spermine (SPM) increase up to 72 h during drought and up to 48 h during salinity stress before their precipitable drop thereafter. Thus, tomato plants maintain survivability to drought as well as salinity stress for up to 3 and 2 days, respectively. Independent multivariant analyses of drought and salinity stress kinetic data separately showed a closer association with levels of free, conjugated, and bound forms of SPD and SPM, but not with free or bound PUT. However, combined multivariant analyses showed a closer association of free SPD, conjugated SPD, and bound SPD with both stresses; SPD-bound and SPM conjugated with drought; and free SPM and conjugated PUT with salinity stress, respectively. PA biosynthesis genes, ARG1, SPDS1, and SAMDc3, segregated with drought and SPDS2 with salinity stress. PA catabolic genes CuAO4-like and PAO4 were associated with drought and salinity stresses, respectively, suggesting differential involvement of PA biosynthesis and catabolic genes in drought and salinity stresses. Pearson correlation indicated mostly positive correlations between the levels of free, conjugated, and bound forms of PUT, SPD, and SPM under drought and salinity stress. However, negative correlations were mostly seen between the levels of various forms of the PAs and their biosynthesis/catabolic genes. Levels of different PA forms had a twofold higher negative correlation during drought as compared to salinity stress (66 vs. 32) and with transcript levels of PA biosynthesis and catabolic genes. Transcripts of light-harvesting chlorophyll a/b-binding genes were generally positively associated with different forms of PAs but negatively to carbon flow genes. Most of the PA biosynthesis genes were coordinately regulated under both stresses. Collectively, these results indicate that PAs are distinctly regulated under drought and salinity stress with different but specific homologs of PA biosynthesis and catabolic genes contributing to the accumulation of free, conjugated, and bound forms of PAs.

Keywords:  drought; polyamine; putrescine (PUT); salt; spermidine (SPD); spermine (SPM); tomato

DOI:  https://doi.org/10.3389/fpls.2021.743568
FEBS J. 2021 Nov 06.

Structure of Pseudomonas aeruginosa spermidine dehydrogenase: a polyamine oxidase with a novel heme-binding fold.

Shiyou Che, Yakun Liang, Yujing Chen, Wenyue Wu, Ruihua Liu, Qionglin Zhang, Mark Bartlam.

  The opportunistic pathogen Pseudomonas aeruginosa can utilize polyamines (including putrescine, cadaverine, 4-aminobutyrate, spermidine and spermine) as its sole source of carbon and nitrogen. Spermidine dehydrogenase (SpdH) is a component of one of the two polyamine utilization pathways identified in P. aeruginosa, but little is known about its structure and function. Here, we report the first crystal structure of SpdH from P. aeruginosa to 1.85 Å resolution. The resulting core structure confirms that SpdH belongs to the polyamine oxidase (PAO) family with flavin-binding and substrate-binding domains. A unique N-terminal extension wraps around the flavin-binding domain of SpdH and is required for heme binding, placing a heme cofactor in close proximity to the FAD cofactor. Structural and mutational analysis reveals that residues in the putative active site at the re side of the FAD isoalloxazine ring form part of the catalytic machinery. PaSpdH features an unusual active site and lacks the conserved lysine that forms part of a lysine-water-flavin N5 atom interaction in other PAO enzymes characterized to date. Mutational analysis further confirms that heme is required for catalytic activity. This work provides an important starting point for understanding the role of SpdH, which occurs universally in P. aeruginosa strains, in polyamine metabolism.

Keywords:   Pseudomonas aeruginosa ; crystal structure; flavoprotein; heme; monoamine oxidase; polyamine metabolism; spermidine dehydrogenase

DOI:  https://doi.org/10.1111/febs.16264
J Bacteriol. 2021 Nov 01. JB0029721

Putrescine and its metabolic precursor arginine promote biofilm and c-di-GMP synthesis in Pseudomonas aeruginosa.

Zhexian Liu, Sarzana S Hossain, Zayda Morales Moreira, Cara H Haney.

Pseudomonas aeruginosa, an opportunistic bacterial pathogen can synthesize and catabolize a number of small cationic molecules known as polyamines. In several clades of bacteria polyamines regulate biofilm formation, a lifestyle-switching process that confers resistance to environmental stress. The polyamine putrescine and its biosynthetic precursors, L-arginine and agmatine, promote biofilm formation in Pseudomonas spp. However, it remains unclear whether the effect is a direct effect of polyamines or through a metabolic derivative. Here we used a genetic approach to demonstrate that putrescine accumulation, either through disruption of the spermidine biosynthesis pathway or the catabolic putrescine aminotransferase pathway, promoted biofilm formation in P. aeruginosa. Consistent with this observation, exogenous putrescine robustly induced biofilm formation in P. aeruginosa that was dependent on putrescine uptake and biosynthesis pathways. Additionally, we show that L-arginine, the biosynthetic precursor of putrescine, also promoted biofilm formation, but via a mechanism independent of putrescine or agmatine conversion. We found that both putrescine and L-arginine induced a significant increase in the intracellular level of bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP) (c-di-GMP), a bacterial second messenger widely found in Proteobacteria that upregulates biofilm formation. Collectively these data show that putrescine and its metabolic precursor arginine promote biofilm and c-di-GMP synthesis in P. aeruginosa. Importance: Biofilm formation allows bacteria to physically attach to a surface, confers tolerance to antimicrobial agents, and promotes resistance to host immune responses. As a result, regulation of biofilm is often crucial for bacterial pathogens to establish chronic infections. A primary mechanism of biofilm promotion in bacteria is the molecule c-di-GMP, which promotes biofilm formation. The level of c-di-GMP is tightly regulated by bacterial enzymes. In this study, we found that putrescine, a small molecule ubiquitously found in eukaryotic cells, robustly enhances P. aeruginosa biofilm and c-di-GMP. We propose that P. aeruginosa may sense putrescine as a host-associated signal that triggers a lifestyle switching that favors chronic infection.

DOI: https://doi.org/10.1128/JB.00297-21
Front Plant Sci. 2021 ;12 701538

Exogenous Spermidine Priming Mitigates the Osmotic Damage in Germinating Seeds of Leymus chinensis Under Salt-Alkali Stress.

Chen Hongna, Shi Junmei, Tao Leyuan, Han Xiaori, Lin Guolin, Cheng Xianguo.

  Spermidine (Spd) is known to protect macromolecules involved in physiological and biochemical processes in plants. However, it is possible that Spd also plays an osmotic regulatory role in promoting the seed germination of Leymus chinensis (L. chinensis) under salt-alkali stress. To investigate this further, seeds of L. chinensis were soaked in Spd solution or distilled water, and a culture experiment was performed by sowing the soaked seeds in saline-alkaline soils. The data showed that the Spd priming resulted in an increase of more than 50% in soluble sugar content and an increase of more than 30% in proline content in the germinating seeds. In addition, the Spd priming resulted in an increase of more than 30% in catalase activity and an increase of more than 25% in peroxidase activity in the germinating seeds and effectively mitigated the oxidative damage to the plasma membrane in the germinating seeds under salt-alkali stress. Moreover, the Spd priming of seeds affected the accumulation of polyamine (PA) and maintained the activities of macromolecules involved in physiological metabolism in germinating seeds exposed to salt-alkali stress. Furthermore, the Spd priming treatment increased the hydrogen peroxide (H2O2) level to more than 30% and the Ca2+ concentration to more than 20% in the germinating seeds, thus breaking the dormancy induction pathways in L. chinensis seeds through beneficial hormone enrichment. This study provides an insight into the Spd-mediated regulation pathway during exogenous Spd priming of L. chinensis seeds, which mitigates osmotic and oxidative damage and maintains the integrality of the cell lipid membrane. Thus, exogenous Spd priming increases PA oxidase activity and maintains the accumulation of H2O2. We found that the H2O2 beneficially affected the balance of Ca2+ and hormones, promoting the vigor and germination of L. chinensis in response to salt-alkali stress.

Keywords:  Leymus chinensis; germination; polyamine oxidase; salt-alkali stress; spermidine

DOI:  https://doi.org/10.3389/fpls.2021.701538
Cell Rep. 2021 Nov 02. pii: S2211-1247(21)01418-2. [Epub ahead of print]37(5): 109945

The endocytic pathway taken by cationic substances requires Rab14 but not Rab5 and Rab7.

Evgeniya Trofimenko, Yuta Homma, Mitsunori Fukuda, Christian Widmann.

  Endocytosis and endosome dynamics are controlled by proteins of the small GTPase Rab family. Besides possible recycling routes to the plasma membrane and various organelles, previously described endocytic pathways (e.g., clathrin-mediated endocytosis, macropinocytosis, CLIC/GEEC pathway) all appear to funnel the endocytosed material to Rab5-positive early endosomes that then mature into Rab7-positive late endosomes/lysosomes. By studying the uptake of a series of cell-penetrating peptides (CPPs), we identify an endocytic pathway that moves material to nonacidic Lamp1-positive late endosomes. Trafficking via this endocytic route is fully independent of Rab5 and Rab7 but requires the Rab14 protein. The pathway taken by CPPs differs from the conventional Rab5-dependent endocytosis at the stage of vesicle formation already, as it is not affected by a series of compounds that inhibit macropinocytosis or clathrin-mediated endocytosis. The Rab14-dependent pathway is also used by physiological cationic molecules such as polyamines and homeodomains found in homeoproteins.

Keywords:  Lamp1; Rab14; Rab5; Rab7; cell-penetrating peptides; endocytosis; endosomes; homeodomains; homeoproteins; polyamines

DOI:  https://doi.org/10.1016/j.celrep.2021.109945