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Molecular Biology of the Cell

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https://www.readbyqxmd.com/read/28515144/transient-activation-of-fission-yeast-ampk-is-required-for-cell-proliferation-during-osmotic-stress
#1
Katherine L Schutt, James B Moseley
The heterotrimeric kinase AMPK acts as an energy sensor to coordinate cell metabolism with environmental status in yeast through humans. Low intracellular ATP leads to AMPK activation through phosphorylation of the activation loop within the catalytic subunit. Other environmental stresses also activate AMPK, but it is unclear if cellular energy status affects AMPK activation under these conditions. Fission yeast AMPK catalytic subunit Ssp2 is phosphorylated at Thr189 by the upstream kinase Ssp1 in low glucose conditions, similar to other systems...
May 17, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28515143/cyclin-c-influences-the-timing-of-mitosis-in-fission-yeast
#2
Gabor Banyai, Zsolt Szilagyi, Vera Baraznenok, Olga Khorosjutina, Claes M Gustafsson
The multiprotein Mediator complex is required for the regulated transcription of nearly all RNA polymerase II-dependent genes. Mediator contains the Cdk8 regulatory subcomplex, which directs periodic transcription and influences cell cycle progression in fission yeast. We here investigate the role of CycC, the cognate cyclin partner of Cdk8, in cell cycle control. Previous reports suggested that CycC interact with other cellular Cdks, but a fusion of CycC to Cdk8 reported here did not cause any obvious cell cycle phenotypes...
May 17, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28515142/flow-induced-endothelial-cell-alignment-requires-the-rhogef-trio-as-a-scaffold-protein-to-polarize-active-rac1-distribution
#3
Jeffrey Kroon, Niels Heemskerk, Martin J T Kalsbeek, Vivian de Waard, Jos van Rijssel, Jaap D van Buul
Endothelial cells line the lumen of the vessel wall and are exposed to flow. In linear parts of the vessel, the endothelial cells experience laminar flow, resulting in endothelial cell alignment in the direction of flow, thereby protecting the vessel wall from inflammation and permeability. In order for endothelial cells to align, they undergo rapid remodeling of the actin cytoskeleton by local activation of the small GTPase Rac1. However, if sustained and local activation of Rac1 is required for long-term flow-induced cell alignment is not clear...
May 17, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28495800/mechanically-patterned-neuromuscular-junctions-in-a-dish-have-improved-functional-maturation
#4
Cassandra L Happe, Kevin P Tenerelli, Anastasia K Gromova, Frederic Kolb, Adam J Engler
Motor neuron (MN) diseases are progressive disorders resulting from degeneration of neuromuscular junctions (NMJs), which form the connection between MNs and muscle fibers. NMJ-in-a-dish models have been developed to examine human MN-associated dysfunction with disease; however such co-culture models have randomly oriented myotubes with immature synapses that contract asynchronously. Mechanically patterned (MP) extracellular matrix with alternating soft and stiff stripes improve current NMJ-in-a-dish models by inducing both mouse and human myoblast durotaxis to stripes where they aligned, differentiated, and fused into patterned myotubes...
May 11, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28495799/studies-on-truncating-mutations-of-spast-associated-with-hereditary-spastic-paraplegia-indicate-greater-accumulation-and-toxicity-of-the-m1-isoform-of-spastin
#5
Joanna M Solowska, Anand N Rao, Peter W Baas
The SPAST gene, which produces two isoforms (M1 and M87) of the microtubule-severing protein called spastin, is the chief gene mutated in hereditary spastic paraplegia. Haploinsufficiency is a popular explanation for the disease, in part because most of the over 200 pathogenic mutations of the gene are truncating, and expected to produce only vanishingly small amounts of shortened proteins. Here we studied two such mutations, N184X and S245X, and our results suggest another possibility. We found that the truncated M1 proteins can accumulate to notably higher levels than their truncated M87 or wild-type counterparts...
May 11, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28495798/a-role-for-sar1-and-arf1-gtpases-during-golgi-biogenesis-in-the-protozoan-parasite-trypanosoma-brucei
#6
Sevil Yavuz, Graham Warren
A single Golgi stack is duplicated and partitioned into two daughter cells during the cell cycle of the protozoan parasite Trypanosoma brucei The source of components required to generate the new Golgi and the mechanism by which it forms is poorly understood. Using photoactivatable GFP we show that the existing Golgi supplies components directly to the newly-forming Golgi, both in intact and semi-permeabilized cells. The movement of a putative glycosyltransferase, GntB, requires the Sar1 and ARF1 GTPases in intact cells...
May 11, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28495797/ixazomib-enhances-parathyroid-hormone-pth-induced-%C3%AE-catenin-t-cell-factor-tcf-signaling-by-dissociating-%C3%AE-catenin-from-the-pth-receptor
#7
Yanmei Yang, Hong Lei, Ya-Wei Qiang, Bin Wang
The anabolic action of PTH in bone is mostly mediated by cAMP/PKA and Wnt-independent activation of β-catenin/T-cell factor (TCF) signaling. β-catenin switches the PTH receptor (PTHR) signaling from cAMP/PKA to PLC/PKC activation by binding to the PTHR. Ixazomib (Izb) is the first orally administered proteasome inhibitor that was approved recently for the treatment of multiple myeloma in part through inhibition of pathologic bone destruction. Proteasome inhibitors were reported to stabilize β-catenin by ubiquitin-proteasome pathway...
May 11, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28495796/phosphorylation-of-nhe3-s-719-regulates-nhe3-activity-through-the-formation-of-multiple-signaling-complexes
#8
Rafiquel Sarker, Boyoung Cha, Olga Kovbasnjuk, Robert Cole, Sandra Gabelli, Chung Ming Tse, Mark Donowitz
Casein kinase 2 (CK2) binds to the NHE3 C-terminus and constitutively phosphorylates a down-stream site (S719) that accounts for 40% of basal NHE3 activity. The role of CK2 in regulation of NHE3 activity in polarized Caco-2/bbe cells was further examined by mutation of NHE3-S(719) to A (not phosphorylated) or D (phosphomimetic). NHE3-S719A but not -S719D had multiple changes in NHE3 activity consisting of a) reduced basal NHE3 activity: specifically, inhibition of the PI3-K/ AKT-dependent component. b) reduced acute stimulation of NHE3 activity by LPA/LPA5R stimulation...
May 11, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28495795/the-desmoplakin-intermediate-filament-linkage-regulates-cell-mechanics
#9
Joshua A Broussard, Ruiguo Yang, Changjin Huang, S Shiva P Nathamgari, Allison M Beese, Lisa M Godsel, Sherry Lee, Fan Zhou, Nathan J Sniadecki, Kathleen J Green, Horacio D Espinosa
The translation of mechanical forces into biochemical signals plays a central role in guiding normal physiological processes during tissue development and homeostasis. Interfering with this process contributes to cardiovascular disease, cancer progression, and inherited disorders. The actin-based cytoskeleton and its associated adherens junctions are well-established contributors to mechanosensing and transduction machinery; however, the role of the desmosome/intermediate filament network is poorly understood in this context...
May 11, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28468979/increased-lateral-microtubule-contact-at-the-cell-cortex-is-sufficient-to-drive-mammalian-spindle-elongation
#10
Joshua Guild, Miriam B Ginzberg, Christina L Hueschen, Timothy J Mitchison, Sophie Dumont
The spindle is a dynamic structure that changes its architecture and size in response to biochemical and physical cues. For example, a simple physical change, cell confinement, can trigger centrosome separation and increase spindle steady-state length at metaphase. How this occurs is not understood, and is the question we pose here. We find that metaphase and anaphase spindles elongate at the same rate when confined, suggesting that similar elongation forces can be generated independent of biochemical and spindle structural differences...
May 3, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28468978/regulation-of-circular-dorsal-ruffles-macropinocytosis-and-cell-migration-by-rhog-and-its-exchange-factor-trio
#11
Alejandra Valdivia, Silvia M Goicoechea, Sahezeel Awadia, Ashtyn Zinn, Rafael Garcia-Mata
Circular dorsal ruffles (CDRs) are actin-rich structures that form on the dorsal surface of many mammalian cells in response to growth factor stimulation. CDRs represent a unique type of structure that forms transiently and only once upon stimulation. The formation of CDRs involves a drastic rearrangement of the cytoskeleton, which is regulated by the Rho family of GTPases. So far, only Rac1 has been consistently associated with CDR formation, whereas the role of other GTPases in this process is either lacking or inconclusive...
May 3, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28468977/mechanical-regulation-of-cardiac-fibroblast-pro-fibrotic-phenotypes
#12
Kate M Herum, Jonas Choppe, Aditya Kumar, Adam J Engler, Andrew D McCulloch
Cardiac fibrosis is a serious condition currently lacking effective treatments. It occurs as a result of cardiac fibroblast activation and differentiation into myofibroblasts, characterized by proliferation, extracellular matrix production and stiffening, and contraction due to the expression of smooth muscle α-actin. The mechanical properties of myocardium change regionally and over time after myocardial infarction. Although mechanical cues are known to activate cardiac fibroblasts, it is unclear which specific mechanical stimuli regulate which specific phenotypic trait; thus we investigated these relationships using three in-vitro models of cardiac fibroblast mechanical activation and found that: 1) Paracrine signaling from stretched cardiomyocytes induces cardiac fibroblast proliferation under mechanical conditions similar to those of the infarct border region; 2) Direct stretch of cardiac fibroblasts mimicking the mechanical environment of the infarct region induces a synthetic phenotype with elevated extracellular matrix production; 3) Progressive matrix stiffening, modeling the mechanical effects of infarct scar maturation, causes smooth muscle α-actin fiber formation, upregulation of collagen I and downregulation of collagen III...
May 3, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28468976/effects-of-substrate-stiffness-and-actomyosin-contractility-on-coupling-between-force-transmission-and-vinculin-paxillin-recruitment-at-single-focal-adhesions
#13
Dennis W Zhou, Ted T Lee, Shinuo Weng, Jianping Fu, Andrés J García
Focal adhesions (FAs) regulate force transfer between the cytoskeleton and ECM-integrin complexes. We previously showed that vinculin regulates force transmission at FAs. Vinculin residence time in FAs correlated with applied force, supporting a mechanosensitive model in which forces stabilize vinculin's active conformation to promote force transfer. In the present study, we examined the relationship between traction force and vinculin-paxillin localization to single FAs in the context of substrate stiffness and actomyosin contractility...
May 3, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28450455/maars-a-novel-high-content-acquisition-software-for-the-analysis-of-mitotic-defects-in-fission-yeast
#14
Tong Li, Hadrien Mary, Marie Grosjean, Jonathan Fouchard, Simon Cabello, Céline Reyes, Sylvie Tournier, Yannick Gachet
Faithful segregation of chromosomes during cell division relies on multiple processes such as chromosome attachment or correct spindle positioning. Yet, mitotic progression is defined by multiple parameters, which need to be quantitatively evaluated. To study the spatio-temporal control of mitotic progression, we developed a High-Content Analysis (HCA) approach, which combines automated fluorescence microscopy with real time quantitative image analysis and allows the unbiased acquisition of multi-parametric data at the single cell level for hundreds of cells simultaneously...
April 27, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28450454/sortilin-and-retromer-mediate-retrograde-transport-of-glut4-in-3t3-l1-adipocytes
#15
Xiang Pan, Nava Zaarur, Maneet Singh, Peter Morin, Konstantin V Kandror
Sortilin is a multi-ligand sorting receptor responsible for the anterograde transport of lysosomal enzymes and substrates. Here, we demonstrate that sortilin is also involved in the retrograde protein traffic. In cultured 3T3-L1 adipocytes, sortilin together with retromer rescues Glut4 from the degradation in lysosomes and retrieves it to the TGN where the insulin-responsive vesicles are formed. Mechanistically, the luminal Vps10p domain of sortilin interacts with the first luminal loop of Glut4, while the cytoplasmic tail of sortilin binds to retromer...
April 27, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28450453/microtubule-dynamics-drive-enhanced-chromatin-motion-and-mobilize-telomeres-in-response-to-dna-damage
#16
Josh Lawrimore, Timothy M Barry, Raymond M Barry, Alyssa C York, Diana M Cook, Kristen Akialis, Jolien Tyler, Paula Vasquez, Elaine Yeh, Kerry Bloom
Chromatin exhibits increased mobility upon DNA damage, but the biophysical basis for this behavior remains unknown. To explore the mechanisms that drive DNA damage-induced chromosome mobility, we employ single-particle tracking of tagged chromosomal loci during interphase in live yeast cells together with polymer models of chromatin chains. Telomeres become mobilized from sites on the nuclear envelope and the pericentromere expands following exposure to DNA damaging agents. The magnitude of chromatin mobility induced by a single double strand break requires active microtubule function...
April 27, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28450452/nanonet-force-microscopy-for-measuring-forces-in-single-smooth-muscle-cells-of-human-aorta
#17
Alexander Hall, Patrick Chan, Kevin Sheets, Matthew Apperson, Christopher Delaughter, Thomas G Gleason, Julie A Phillippi, Amrinder Nain
A number of innovative methods exist to measure cell-matrix adhesive forces, which have yet to accurately describe and quantify the intricate interplay of a cell and its fibrous extracellular matrix (ECM). In cardiovascular pathologies, such as aortic aneurysm, new knowledge on the involvement of cell-matrix forces could lead to elucidation of disease mechanisms. To better understand this dynamics we measured primary human aortic single smooth muscle cell (SMC) forces using nanonet force microscopy (NFM) in both inside-out (I-O intrinsic contractility) and outside-in (O-I external perturbation) modes...
April 27, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28450451/a-novel-munc13-4-s100a10-annexin-a2-complex-promotes-weibel-palade-body-exocytosis-in-endothelial-cells
#18
Tarek Chehab, Nina Criado Santos, Anna Holthenrich, Sophia N Koerdt, Jennifer Disse, Christian Schuberth, Ali Reza Nazmi, Maaike Neeft, Henriette Koch, Kwun Nok M Man, Sonja M Wojcik, Thomas F J Martin, Peter van der Sluijs, Nils Brose, Volker Gerke
Endothelial cells respond to blood vessel injury by the acute release of pro-coagulant von-Willebrand factor (VWF) that is stored in unique secretory granules called Weibel-Palade bodies (WPBs). Stimulated WPB exocytosis critically depends on the proper recruitment of WPBs to the plasma membrane but factors involved in WPB-plasma membrane tethering are not known. Here we identify Munc13-4, a protein mutated in familial hemophagocytic lymphohistiocytosis 3 (FHL3), as a WPB tethering factor. Munc13-4 promotes histamine-evoked WPB exocytosis, it is present on WPBs, and secretagogue stimulation triggers an increased recruitment of Munc13-4 to WPBs and a clustering of Munc13-4 at sites of WPB-plasma membrane contact...
April 27, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28428259/rabl2-interacts-with-the-ift-b-complex-and-cep19-and-participates-in-ciliary-assembly
#19
Yuya Nishijima, Yohei Hagiya, Tomohiro Kubo, Ryota Takei, Yohei Katoh, Kazuhisa Nakayama
Proteins localized to the basal body and the centrosome play crucial roles in ciliary assembly and functions. Although RABL2 and CEP19 are conserved in ciliated organisms and have been implicated in ciliary/flagellar functions, their roles were poorly understood. We here showed that RABL2 interacts with CEP19 and is recruited to the mother centriole and basal body in a CEP19-dependent manner, and that CEP19 is recruited to the centriole probably via its binding to the centrosomal protein FGFR1OP. Disruption of the RABL2 gene in Chlamydomonas reinhardtii resulted in the non-flagellated phenotype, suggesting a crucial role of RABL2 in ciliary/flagellar assembly...
April 20, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28428258/deletion-of-inositol-requiring-enzyme-1%C3%AE-in-podocytes-disrupts-glomerular-capillary-integrity-and-autophagy
#20
Daniel Robert Kaufman, Joan Papillon, Louise Larose, Takao Iwawaki, Andrey V Cybulsky
Inositol-requiring enzyme-1α (IRE1α) is an endoplasmic reticulum (ER)-transmembrane endoribonuclease-kinase, which plays an essential function in extraembryonic tissues during normal development, and is activated during ER stress. To address the functional role of IRE1α in glomerular podocytes, we produced podocyte-specific IRE1α deletion mice. In male mice, deletion of IRE1α in podocytes resulted in albuminuria beginning at 5 months of age, and worsening with time. Electron microscopy revealed focal podocyte foot process effacement in 9-month old male IRE1α deletion mice, as well as microvillous transformation of podocyte plasma membranes...
April 20, 2017: Molecular Biology of the Cell
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