bims-micesi 2021-12-19 papers

bims-micesi

Biomed News

on Mitotic cell signalling

Issue of 2021–12–19
nine papers selected by
Valentina Piano, Max Planck Institute of Molecular Physiology

Septin Assembly and Remodeling at the Cell Division Site During the Cell Cycle.
More than Two Populations of Microtubules Comprise the Dynamic Mitotic Spindle.
A review on synthetic chalcone derivatives as tubulin polymerisation inhibitors.
The centriolar tubulin code.
Maternal Smc3 protects the integrity of the zygotic genome through DNA replication and mitosis.
The effect of danusertib, an Aurora kinase inhibitor, onto the cytotoxicity, cell cycle and apoptosis in pancreatic ductal adenocarcinoma cells.
Lattice defects induced by microtubule-stabilizing agents exert a long-range effect on microtubule growth by promoting catastrophes.
Photoswitchable epothilone-based microtubule stabilisers allow GFP-imaging-compatible, optical control over the microtubule cytoskeleton.
Aurora kinase A induces migration and invasion by inducing epithelial-to-mesenchymal transition in colon cancer cells.

Front Cell Dev Biol. 2021 ;9 793920

Septin Assembly and Remodeling at the Cell Division Site During the Cell Cycle.

Joseph Marquardt, Xi Chen, Erfei Bi.

  The septin family of proteins can assemble into filaments that further organize into different higher order structures to perform a variety of different functions in different cell types and organisms. In the budding yeast Saccharomyces cerevisiae, the septins localize to the presumptive bud site as a cortical ring prior to bud emergence, expand into an hourglass at the bud neck (cell division site) during bud growth, and finally "split" into a double ring sandwiching the cell division machinery during cytokinesis. While much work has been done to understand the functions and molecular makeups of these structures, the mechanisms underlying the transitions from one structure to another have largely remained elusive. Recent studies involving advanced imaging and in vitro reconstitution have begun to reveal the vast complexity involved in the regulation of these structural transitions, which defines the focus of discussion in this mini-review.

Keywords:  Bud3; Bud4; Elm1; RhoGEF; anillin; septin-associated proteins; septins

DOI:  https://doi.org/10.3389/fcell.2021.793920
J Cell Sci. 2021 Dec 15. pii: jcs.258745. [Epub ahead of print]

More than Two Populations of Microtubules Comprise the Dynamic Mitotic Spindle.

Aaron R Tipton, Gary J Gorbsky.

  The microtubules of the mitotic spindle mediate chromosome alignment to the metaphase plate, then sister chromatid segregation to the spindle poles in anaphase. Previous analyses of spindle microtubule kinetics utilizing fluorescence dissipation after photoactivation described two main populations, a slow and a fast turnover population, and these were ascribed to reflect kinetochore versus non-kinetochore microtubules, respectively. Here, we test this categorization by disrupting kinetochores through depletion of the Ndc80 complex. In the absence of functional kinetochores, microtubule dynamics still exhibit slow and fast turnover populations, though the proportion of each population and the timings of turnover are altered. Importantly, the data obtained following Hec1/Ndc80 depletion suggests other sub-populations, in addition to kinetochore microtubules, contribute to the slow turnover population. Further manipulation of spindle microtubules revealed a complex landscape. For example, while Aurora B kinase functions to destabilize kinetochore bound microtubules it may also stabilize certain slow turnover, non-kinetochore microtubules. Dissection of the dynamics of microtubule populations provides a greater understanding of mitotic spindle kinetics and insight into their roles in facilitating chromosome attachment, movement, and segregation during mitosis.

Keywords:  Cell cycle; Chromosome; Fluorescence; Microtubules; Mitosis; Mitotic spindle

DOI:  https://doi.org/10.1242/jcs.258745
J Enzyme Inhib Med Chem. 2022 Dec;37(1): 9-38

A review on synthetic chalcone derivatives as tubulin polymerisation inhibitors.

Wenjing Liu, Min He, Yongjun Li, Zhiyun Peng, Guangcheng Wang.

  Microtubules play an important role in the process of cell mitosis and can form a spindle in the mitotic prophase of the cell, which can pull chromosomes to the ends of the cell and then divide into two daughter cells to complete the process of mitosis. Tubulin inhibitors suppress cell proliferation by inhibiting microtubule dynamics and disrupting microtubule homeostasis. Thereby inducing a cell cycle arrest at the G2/M phase and interfering with the mitotic process. It has been found that a variety of chalcone derivatives can bind to microtubule proteins and disrupt the dynamic balance of microtubules, inhibit the proliferation of tumour cells, and exert anti-tumour effects. Consequently, a great number of studies have been conducted on chalcone derivatives targeting microtubule proteins. In this review, synthetic or natural chalcone microtubule inhibitors in recent years are described, along with their structure-activity relationship (SAR) for anticancer activity.

Keywords:  Chalcones; anticancer; microtubule; tubulin polymerisation inhibitors

DOI:  https://doi.org/10.1080/14756366.2021.1976772
Semin Cell Dev Biol. 2021 Dec 09. pii: S1084-9521(21)00309-8. [Epub ahead of print]

The centriolar tubulin code.

Paul Guichard, Marine H Laporte, Virginie Hamel.

  Centrioles are microtubule-based cell organelles present in most eukaryotes. They participate in the control of cell division as part of the centrosome, the major microtubule-organizing center of the cell, and are also essential for the formation of primary and motile cilia. During centriole assembly as well as across its lifetime, centriolar tubulin display marks defined by post-translational modifications (PTMs), such as glutamylation or acetylation. To date, the functions of these PTMs at centrioles are not well understood, although pioneering experiments suggest a role in the stability of this organelle. Here, we review the current knowledge regarding PTMs at centrioles with a particular focus on a possible link between these modifications and centriole's architecture, and propose possible hypothesis regarding centriolar tubulin PTMs's function.

Keywords:  Basal bodies; Centrioles; Cryo-EM; Expansion microscopy; PTMs; Tubulin

DOI:  https://doi.org/10.1016/j.semcdb.2021.12.001
Development. 2021 Dec 15. pii: dev.199800. [Epub ahead of print]

Maternal Smc3 protects the integrity of the zygotic genome through DNA replication and mitosis.

Wei-Ting Yueh, Vijay Pratap Singh, Jennifer L Gerton.

  Aneuploidy is frequently observed in oocytes and early embryos, begging the question of how genome integrity is monitored and preserved during this critical period. SMC3 is a subunit of the cohesin complex that supports genome integrity, but its role in maintaining the genome in this window of mammalian development is unknown. We discovered that although depletion of Smc3 following meiotic S phase in mouse oocytes allowed accurate meiotic chromosome segregation, adult females were infertile. We provide evidence that DNA lesions accumulated following S phase in SMC3-deficient zygotes, followed by mitosis with lagging chromosomes, elongated spindles, micronuclei, and arrest at the 2-cell stage. Remarkably, although centromeric cohesion was defective, the dosage of SMC3 was sufficient to enable embryogenesis in juvenile mutant females. Our findings suggest that despite previous reports of aneuploidy in early embryos, chromosome missegregation in zygotes halts embryogenesis at the 2-cell stage. Smc3 is a maternal gene with essential functions in repair of spontaneous damage associated with DNA replication and subsequent chromosome segregation in zygotes, making cohesin a key protector of the zygotic genome.

Keywords:  Chromosome segregation; Cohesin; Developmental competence; Juvenile; Maternal effect gene; Micronuclei; Mouse; SMC3; Spontaneous DNA damage; Zygote

DOI:  https://doi.org/10.1242/dev.199800
J Cancer Res Ther. 2021 Oct-Dec;17(6):17(6): 1419-1424

The effect of danusertib, an Aurora kinase inhibitor, onto the cytotoxicity, cell cycle and apoptosis in pancreatic ductal adenocarcinoma cells.

Ismail Ayberk Kirbiyik, Ahmet Ata Ozcimen.

   Background: Pancreatic cancer is the second type of cancer that causes the most death among the digestive system cancers. Difficulties in early diagnosis and rapidly progressing to advanced stages are most common in high mortality rate of pancreatic carcinoma. The mutation of Bcr-Abl tyrosine kinase and mitotic kinases (such as Aurora kinases), which are involved in the cell cycle, plays an important role in the progression of cancer. Enzymes belonging to Aurora kinase family (-A, -B, -C) have been reported to play a major role in cancer progression, invasion and metastasis. Therefore, the purpose of this study, investigate of the effect of danusertib, an Aurora kinase inhibitor, onto cytotoxicity, apoptosis and cell cycle in human pancreatic carcinoma CFPAC-1 cells.
Materials and Methods: For determining the IC50 value, the 20,000 cells were seeded in E-plate 16 wells in a real-time cell analyzer and various concentrations of danusertib (1-10,000 nM) were applied onto CFPAC-1 cells incubated in IMDM medium. Cell index demonstrated that the proliferation of fraction cells was measured in real time. On the other hand, cell apoptosis and cell cycle arrest test were stained with Annexin V-APC/PI and DNA-cell cycle PI staining respectively by using flow cytometry.
Results: The IC50 value was found to be approximately 400 nM. Danusertib at this concentration induced apoptosis in CFPAC-1 cells (%14,8 at 24 hours; %21,3 at 48 hours). Furthermore, in the cells treated with danusertib, 31.77% and 11.05% were arrested in the S and G2 phases, respectively.
Conclusions: Aurora kinase inhibitor danusertib induced a significant effect of cytotoxic, apoptotic and cell cycle arrest in CFPAC-1 ductal adenocarcinoma cells. Therefore, it may be a potential alternative to the treatment of pancreatic cancers.

Keywords:  Apoptosis; Aurora kinase; cell cycle; cystic fibrosis pancreatic adenocarcinoma cell line-1; danusertib

DOI:  https://doi.org/10.4103/jcrt.JCRT_827_19
Proc Natl Acad Sci U S A. 2021 Dec 21. pii: e2112261118. [Epub ahead of print]118(51):

Lattice defects induced by microtubule-stabilizing agents exert a long-range effect on microtubule growth by promoting catastrophes.

Ankit Rai, Tianyang Liu, Eugene A Katrukha, Juan Estévez-Gallego, Szymon W Manka, Ian Paterson, J Fernando Díaz, Lukas C Kapitein, Carolyn A Moores, Anna Akhmanova.

  Microtubules are dynamic cytoskeletal polymers that spontaneously switch between phases of growth and shrinkage. The probability of transitioning from growth to shrinkage, termed catastrophe, increases with microtubule age, but the underlying mechanisms are poorly understood. Here, we set out to test whether microtubule lattice defects formed during polymerization can affect growth at the plus end. To generate microtubules with lattice defects, we used microtubule-stabilizing agents that promote formation of polymers with different protofilament numbers. By employing different agents during nucleation of stable microtubule seeds and the subsequent polymerization phase, we could reproducibly induce switches in protofilament number and induce stable lattice defects. Such drug-induced defects led to frequent catastrophes, which were not observed when microtubules were grown in the same conditions but without a protofilament number mismatch. Microtubule severing at the site of the defect was sufficient to suppress catastrophes. We conclude that structural defects within the microtubule lattice can exert effects that can propagate over long distances and affect the dynamic state of the microtubule end.

Keywords:  Taxol; in vitro reconstitution; microtubule; photoablation; protofilament

DOI:  https://doi.org/10.1073/pnas.2112261118
Angew Chem Int Ed Engl. 2021 Dec 13.

Photoswitchable epothilone-based microtubule stabilisers allow GFP-imaging-compatible, optical control over the microtubule cytoskeleton.

Li Gao, Joyce C M Meiring, Constanze Heise, Ankit Rai, Adrian Müller-Deku, Anna Akhmanova, Julia Thorn-Seshold, Oliver Thorn-Seshold.

  Optical methods to modulate microtubule dynamics have promise for reaching the micron- and millisecond-scale resolution needed to decrypt the roles of the cytoskeleton in biology. However, optical microtubule stabilisers are under-developed. We introduce "STEpos" as GFP-orthogonal, light-responsive epothilone-based microtubule stabilisers. They use a novel styrylthiazole photoswitch in a design to modulate hydrogen-bonding and steric effects that control epothilone potency. STEpos photocontrol microtubule dynamics and cell division with micron- and second-scale spatiotemporal precision. They substantially improve potency, solubility, and ease-of-use compared to previous optical microtubule stabilisers, and the structure-photoswitching-activity relationship insights in this work will guide future optimisations. The STEpo reagents can contribute greatly to high-precision research in cytoskeleton biophysics, cargo transport, cell motility, cell division, development, and neuroscience.

Keywords:  bioorganic chemistry; microtubule inhibitor; natural products; photopharmacology; photoswitch

DOI:  https://doi.org/10.1002/anie.202114614
BMB Rep. 2021 Dec 14. pii: 5511. [Epub ahead of print]

Aurora kinase A induces migration and invasion by inducing epithelial-to-mesenchymal transition in colon cancer cells.

On-Yu Hong, Sang Yull Kang, Eun-Mi Noh, Hong-Nu Yu, Hye-Yeon Jang, Seong Hoon Kim, Jingyu Hong, Eun Yong Chung, Jong-Suk Kim.

Aurora kinase is a family of serine/threonine kinases intimately associated with mitotic progression and the development of human cancers. Studies have shown that aurora kinases are important for the protein kinase C (PKC)-induced invasion of colon cancer cells. Recent studies have shown that aurora kinase A promotes distant metastasis by inducing epithelial-to-mesenchymal transition (EMT) in colon cancer cells. However, the role of aurora kinase A in colon cancer metastasis remains unclear. In this study, we investigated the effects of aurora kinase A on PKC-induced cell invasion, migration, and EMT in human SW480 colon cancer cells. Treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA) changed the expression levels of EMT markers, increasing α-SMA, vimentin, and MMP-9 expression and decreasing E-cadherin expression, with changes in cell morphology. TPA treatment induced EMT in a PKC-dependent manner. Moreover, the inhibition of aurora kinase A by siRNAs and inhibitors (reversine and VX-680) suppressed TPA-induced cell invasion, migration, and EMT in SW480 human colon cells. Inhibition of aurora kinase A blocked TPA-induced vimentin and MMP-9 expression, and decreased E-cadherin expression. Furthermore, the knockdown of aurora kinase A decreased the transcriptional activity of NF-κB and AP-1 in PKC-stimulated SW480 cells. These findings indicate that aurora kinase A induces migration and invasion by inducing EMT in SW480 colon cancer cells. To the best of our knowledge, this is the first study that showed aurora kinase A is a key molecule in PKC-induced metastasis in colon cancer cells.