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Journal of Cell Science

Xian Wang, Haijiao Liu, Min Zhu, Changhong Cao, Zhensong Xu, Yonit Tsatskis, Kimberly Lau, Chikin Kuok, Tobin Filleter, Helen McNeill, Craig A Simmons, Sevan Hopyan, Yu Sun
Extracellular forces transmitted through the cytoskeleton can deform the cell nucleus. Large nuclear deformation increases the risk of disrupting the nuclear envelope's integrity and causing DNA damage. Mechanical stability of the nucleus defines its capability of maintaining nuclear shape by minimizing nuclear deformation and recovering strain when deformed. Understanding the deformation and recovery behavior of the nucleus requires characterization of nuclear viscoelastic properties. Here, we quantified the decoupled viscoelastic parameters of the cell membrane, cytoskeleton, and the nucleus...
May 18, 2018: Journal of Cell Science
Matthew N J Seaman, Aamir S Mukadam, Sophia Y Breusegem
The retromer complex is a vital component of the endosomal protein sorting machinery being necessary for sorting into both the endosome-to-Golgi retrieval pathway and also the endosome-to-cell-surface recycling pathway. Retromer mediates cargo selection through a trimeric complex comprising VPS35, VPS29 and VPS26 which is recruited to endosomes by binding to Rab7a and Snx3. Retromer function is linked to two distinct neurodegenerative diseases, Parkinson's disease and Alzheimer's disease and modulating retromer function has been proposed as an avenue to explore for a putative therapy in these conditions...
May 18, 2018: Journal of Cell Science
Kiminori Kurashima, Takayuki Sekimoto, Tsukasa Oda, Tsuyoshi Kawabata, Fumio Hanaoka, Takayuki Yamashita
Growth of precancerous and cancer cells relies on their tolerance of oncogene-induced replication stress (RS). Translesion synthesis (TLS) plays an essential role in cellular tolerance of various types of RS and bypasses replication barriers by employing specialized polymerases. However, limited information is available about the role of TLS polymerases in oncogene-induced RS. Here, we report that Polη, a Y-family TLS polymerase, promotes cellular tolerance of Myc-induced RS. Polη was recruited to Myc-induced RS sites, and Polη depletion enhanced the Myc-induced slowing and stalling of replication forks and the subsequent generation of double-strand breaks (DSBs)...
May 18, 2018: Journal of Cell Science
Asako Shindo, Anastasia Audrey, Maki Takagishi, Masahide Takahashi, John B Wallingford, Makoto Kinoshita
Wounds in embryos heal rapidly through the contraction of the wound edges. Despite well-recognized significance of the actomyosin purse-string for wound closure, roles for other cytoskeletal components are largely unknown. Here, we report that the septin cytoskeleton cooperates with actomyosin and microtubules to coordinate circumferential contraction of the wound margin and concentric elongation of wound-proximal cells in Xenopus laevis embryos. Microtubules reoriented radially, forming bundles along lateral cell cortices in elongating wound-proximal cells...
May 18, 2018: Journal of Cell Science
Jan Steinkühler, Bartosz Różycki, Cory Alvey, Reinhard Lipowsky, Thomas R Weikl, Rumiana Dimova, Dennis E Discher
Cell-cell interactions that result from membrane proteins binding weakly in trans can cause accumulations in cis that suggest cooperativity and thereby an acute sensitivity to environmental factors. The ubiquitous "marker of self" protein CD47 binds weakly to SIRPα on a macrophage, which leads to accumulation of SIRPα at a phagocytic synapse and ultimately to inhibition of engulfment of 'self' cells - including cancer cells. We reconstituted this macrophage checkpoint with CD47-GFP displayed on giant vesicles generated from plasma membranes and then imaged vesicles adhering to SIRPα immobilized on a surface...
May 18, 2018: Journal of Cell Science
Angela Rey-Gallardo, Hannah Tomlins, Justin Joachim, Izajur Rahman, Phoebe Kitscha, Karen Frudd, Maddy Parsons, Aleksandar Ivetic
Leukocyte transendothelial migration (TEM) is absolutely fundamental to the inflammatory response, involving initial pseudopod protrusion and subsequent polarised migration across inflamed endothelium. Ezrin/radixin/moesin (ERM) proteins are expressed in leukocytes and mediate cell shape change and polarity. The spatio-temporal organisation of ERM with their targets, and their individual contribution to protrusion during TEM, has never been explored. Here we show that blocking PIP2 binding reduces C-terminal phosphorylation of moesin during monocyte TEM, and that on/off cycling of ERM activity is essential for pseudopod protrusion into the subendothelial space...
May 18, 2018: Journal of Cell Science
Ye Zhang, Jian-Feng Chang, Jin Sun, Lu Chen, Xiao-Mei Yang, Huan-Yin Tang, Yuan-Ya Jing, Xuan Kang, Zhi-Min He, Jun-Yu Wu, Hui-Min Wei, Da-Liang Wang, Rong-Gang Xu, Rui-Bao Zhu, Ying Shen, Shi-Yang Zeng, Chen Wang, Kui-Nan Liu, Yong Zhang, Zhi-Ying Mao, Ci-Zhong Jiang, Fang-Lin Sun
Dysregulation of homeostatic balance in di- and tri-methyl H3K27 levels or that caused by mis-sense mutations of histone H3 (H3K27M) was reported to be associated with various types of cancers. In this study, we found that reduction in H3K27me2/3 caused by H3.1K27M, a mutation of H3 variants found in DIPG patients, dramatically attenuated the presence of 53BP1 foci and NHEJ repair capability in HDF cells. H3.1K27M cells showed increased rates of genomic insertions/deletions (In/Dels) and copy number variations (CNVs), as well as augmented p53-dependent apoptotic cells...
May 14, 2018: Journal of Cell Science
Travis J Armiger, Marsha C Lampi, Cynthia A Reinhart-King, Kris N Dahl
Force generation within cells, mediated by motor proteins along cytoskeletal networks, maintains the function of multicellular structures during homeostasis and when generating collective forces. Here, we describe the use of chromatin dynamics to detect cellular force propagation (a technique termed Sensors from IntraNuclear Kinetics (SINK)) and investigate the force response of cells to disruption of the monolayer and changes in substrate stiffness. We find that chromatin dynamics change in a substrate stiffness dependent manner within epithelial monolayers...
May 10, 2018: Journal of Cell Science
Rong Fu, Yi Zhu, Xiaowan Jiang, Yuanbao Li, Ming Zhu, Mengqiu Dong, Zhaohui Huang, Chunxia Wang, Michel Labouesse, Huimin Zhang
Hemidesmosomes are epithelial-specific attachment structures that maintain tissue integrity and resist tension. Despite their importance, how hemidesmosomes are regulated at the post-transcriptional level is poorly understood. C. elegans hemidesmosomes (CeHDs) share similar structure and composition with their mammalian counterparts, making C. elegans an ideal model for studying hemidesmosomes. Here we focus on the transcription regulator CCAR-1, identified in a previous genetic screen searching for enhancers of mutations in the conserved hemidesmosome component, VAB-10A/Plectin...
May 10, 2018: Journal of Cell Science
Natalie E Farrawell, Isabella Lambert-Smith, Kristen Mitchell, Jessie McKenna, Luke McAlary, Prajwal Ciryam, Kara L Vine, Darren N Saunders, Justin J Yerbury
A hallmark of amyotrophic lateral sclerosis (ALS) pathology is the accumulation of ubiquitinated protein inclusions within motor neurons. Recent studies suggest the sequestration of ubiquitin (Ub) into inclusions reduces the availability of free Ub, which is essential for cellular function and survival. However, the dynamics of the Ub landscape in ALS have not yet been described. Here we show that Ub homeostasis is altered in a SOD1 cell model of ALS. Monitoring the distribution of Ub in SOD1-expressing cells, we show that Ub is present at the earliest stages of SOD1 aggregation and cells containing mutant SOD1 aggregates have greater ubiquitin-proteasome system (UPS) dysfunction...
May 10, 2018: Journal of Cell Science
Nivetha Kannan, Vivian W Tang
Actomyosin II contractility in epithelial cell plays an essential role in tension-dependent adhesion strengthening. One key unsettling question is how cellular contraction transmits force to the nascent cell-cell adhesion when there is no stable attachment between the nascent adhesion complex and actin filament. Here, we show that myosin-1c is localized to the lateral membrane of polarized epithelial cells and facilitates the coupling between actin and cell-cell adhesion. Knockdown of myosin-1c compromised the integrity of the lateral membrane, reduced the generation of tension at E-cadherin, decreased the strength of cell-cell cohesion in an epithelial cell monolayer, and prevented force-dependent recruitment of junctional α-actinin-4...
May 10, 2018: Journal of Cell Science
Ellena Hönig, Karina Ringer, Jenny Dewes, Tobias von Mach, Natalia Kamm, Geri Kreitzer, Ralf Jacob
Epithelial cells require a precise intracellular transport and sorting machinery in order to establish and maintain their polarized architecture. This machinery includes beta-galactoside binding galectins for glycoprotein targeting to the apical membrane. Galectin-3 sorts cargo destined for the apical plasma membrane into vesicular carriers. After delivery of cargo to the apical milieu, galectin-3 recycles back into sorting organelles. We analyzed the role of galectin-3 in the polarized distribution of β1-integrin in MDCK cells...
May 10, 2018: Journal of Cell Science
Oddrun Elise Olsen, Meenu Sankar, Samah Elsaadi, Hanne Hella, Glenn Buene, Sagar Ramesh Darvekar, Kristine Misund, Takenobu Katagiri, Petra Knaus, Toril Holien
TGF-β/BMP superfamily ligands require heteromeric complexes of type 1 and 2 receptors for ligand dependent downstream signaling. Activin A, a TGF-β superfamily member, inhibits growth of multiple myeloma cells, but the mechanism is unknown. We aimed to clarify how activins affect myeloma cell survival. Activin A activates the transcription factors SMAD2/3 through the ALK4 type 1 receptor, but may also activate SMAD1/5/8 through mutated variants of the type 1 receptor ALK2. We demonstrate that activin A and B activate SMAD1/5/8 in myeloma cells through endogenous wild type ALK2...
May 8, 2018: Journal of Cell Science
Beryl Augustine, Cheen Fei Chin, Foong May Yeong
Mitotic spindle dynamics are regulated during the cell cycle by microtubule motor proteins. One such protein is Kip2p, a plus-ended motor that regulates the polymerization and stability of cytoplasmic microtubules (cMTs). Kip2p levels are regulated during the cell cycle and its overexpression leads to the formation of hyper-elongated cMTs. To investigate the significance of varying Kip2p levels during the cell cycle and the hyper-elongated cMTs, we overexpressed KIP2 in G1 phase and examined the effects on the separation of spindle pole bodies (SPBs) and chromosome segregation...
May 8, 2018: Journal of Cell Science
María Gracia Gervasi, Xinran Xu, Blanca Carbajal-Gonzalez, Mariano G Buffone, Pablo E Visconti, Diego Krapf
Conception of a new mammalian organism is determined by the fusion of a sperm cell with an oocyte during fertilization. Motility is one of the features of sperm that allows them to succeed in fertilization, and their flagellum is essential for this function. Longitudinally, the flagellum divides into the midpiece, the principal piece, and the end piece. A precise cytoskeletal architecture of the sperm tail is key for the acquisition of fertilization competence. It has been proposed that the actin cytoskeleton plays essential roles in the regulation of sperm motility, however, actin organization in sperm remains elusive...
May 8, 2018: Journal of Cell Science
Zhimin Wang, Floris Bosveld, Yohanns Bellaïche
In epithelial tissue, new cell-cell junctions are formed upon cytokinesis. To understand junction formation during cytokinesis, we explored in Drosophila epithelium, de novo formation of tricellular septate junctions (TCJs). We found that upon midbody formation, the membranes of the two daughter cells and of the neighbouring cells located below the adherens junction (AJ) remain entangled in a 4-cell structure apposed to the midbody. The septate junction protein Discs-Large and components of the TCJ, Gliotactin and Anakonda accumulate in this 4-cell structure...
May 8, 2018: Journal of Cell Science
Alexandre Maucuer, Bénédicte Desforges, Vandana Joshi, Mirela Boca, Dmitry Kretov, Loic Hamon, Ahmed Bouhss, Patrick A Curmi, David Pastré
Liquid-liquid phase separation enables compartmentalization of biomolecules in cells, notably RNA and associated proteins in the nucleus. Besides critical functions in RNA processing, there is a major interest in deciphering the molecular mechanisms of compartmentalization orchestrated by RNA-binding proteins such as TDP-43 and FUS due to their link to neuron diseases. However, tools for probing compartmentalization in cells are lacking. Here we developed a method to analyze the mixing:demixing of two different phases in a cellular context...
May 4, 2018: Journal of Cell Science
María Gabriela Otero, Ivan Fernandez Bessone, Alan Earle Hallberg, Lucas Eneas Cromberg, María Cecilia De Rossi, Trinidad M Saez, Valeria Levi, Angels Almenar-Queralt, Tomás Luis Falzone
Alzheimer Disease (AD) pathology includes the accumulation of poly-ubiquitinated proteins and failures in proteasome-dependent degradation. Whereas the distribution of proteasomes and its role in synaptic function have been studied, whether proteasome activity regulates the axonal transport and metabolism of the amyloid precursor protein (APP), remains elusive. Using live imaging in primary hippocampal neurons, we showed that proteasome inhibition rapidly and severely impairs the axonal transport of APP. Fluorescent cross-correlation analyses and membrane internalization blockage showed that plasma membrane APP do not contribute to transport defects...
May 3, 2018: Journal of Cell Science
Peter G Wolf, Alexander Cuba Ramos, Julia Kenzel, Brigitte Neumann, Olaf Stemmann
The DNA embracing, ring-shaped, multiprotein complex cohesin mediates sister chromatid cohesion and is stepwise displaced in mitosis by Wapl and Separase to facilitate anaphase. Proper regulation of chromosome cohesion throughout meiosis is critical to prevent formation of aneuploid gametes, which are associated with trisomies and infertility in humans. Studying cohesion in meiocytes is complicated by their difficult experimental amenability and the absence of cohesin turnover. Here, we use cultured somatic cells to unravel fundamental aspects of meiotic cohesin...
May 3, 2018: Journal of Cell Science
Yongqiang Wang, Matthew Galli, Alexandra Shade Silver, Wilson Lee, Yushan Song, Yixue Michelle Mei, Carly Bachus, Michael Glogauer, Christopher A McCulloch
Adseverin is an actin binding protein involved in osteoclastogenesis, but its role in inflammation-induced bone loss is not defined. We examined whether IL1β and TNFα regulate adseverin expression to control osteoclastogenesis in mouse primary monocytes and RAW264.7 cells. Adseverin co-localized with subcortical actin filaments and was enriched in fusopods of fusing cells. In precursor cells, adseverin overexpression boosted RANKL-induced multinucleated cell formation. IL1β and TNFα enhanced RANKL-dependent TRAcP activity by 1...
May 3, 2018: Journal of Cell Science
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