scholarly journals NudCL2 regulates cell migration by stabilizing both myosin-9 and LIS1 with Hsp90

2020 ◽  
Vol 11 (7) ◽  
Author(s):  
Wenwen Chen ◽  
Wei Wang ◽  
Xiaoxia Sun ◽  
Shanshan Xie ◽  
Xiaoyang Xu ◽  
...  
Keyword(s):  
Cell Migration ◽  
Single Cell ◽  
Mammalian Cells ◽  
Ectopic Expression ◽  
Heat Shock Protein 90 ◽  
Underlying Mechanism ◽  
Collective Cell Migration ◽  
Interacting Protein ◽  
Protein Levels ◽  
Protein Instability

Abstract Cell migration plays pivotal roles in many biological processes; however, its underlying mechanism remains unclear. Here, we find that NudC-like protein 2 (NudCL2), a cochaperone of heat shock protein 90 (Hsp90), modulates cell migration by stabilizing both myosin-9 and lissencephaly protein 1 (LIS1). Either knockdown or knockout of NudCL2 significantly increases single-cell migration, but has no significant effect on collective cell migration. Immunoprecipitation–mass spectrometry and western blotting analyses reveal that NudCL2 binds to myosin-9 in mammalian cells. Depletion of NudCL2 not only decreases myosin-9 protein levels, but also results in actin disorganization. Ectopic expression of myosin-9 efficiently reverses defects in actin disorganization and single-cell migration in cells depleted of NudCL2. Interestingly, knockdown of myosin-9 increases both single and collective cell migration. Depletion of LIS1, a NudCL2 client protein, suppresses both single and collective cell migration, which exhibits the opposite effect compared with myosin-9 depletion. Co-depletion of myosin-9 and LIS1 promotes single-cell migration, resembling the phenotype caused by NudCL2 depletion. Furthermore, inhibition of Hsp90 ATPase activity also reduces the Hsp90-interacting protein myosin-9 stability and increases single-cell migration. Forced expression of Hsp90 efficiently reverses myosin-9 protein instability and the defects induced by NudCL2 depletion, but not vice versa. Taken together, these data suggest that NudCL2 plays an important role in the precise regulation of cell migration by stabilizing both myosin-9 and LIS1 via Hsp90 pathway.

scholarly journals NudC L279P Mutation Destabilizes Filamin A by Inhibiting the Hsp90 Chaperoning Pathway and Suppresses Cell Migration

2021 ◽  
Vol 9 ◽  
Author(s):  
Min Liu ◽  
Zhangqi Xu ◽  
Cheng Zhang ◽  
Chunxia Yang ◽  
Jiaxing Feng ◽  
...  
Keyword(s):  
Cell Migration ◽  
Protein Stability ◽  
Mammalian Cells ◽  
Regulatory Mechanism ◽  
Ectopic Expression ◽  
Heat Shock Protein 90 ◽  
Muscle Actin ◽  
Filamin A ◽  
Evolutionarily Conserved ◽  
Functional Defects

Filamin A, the first discovered non-muscle actin filament cross-linking protein, plays a crucial role in regulating cell migration that participates in diverse cellular and developmental processes. However, the regulatory mechanism of filamin A stability remains unclear. Here, we find that nuclear distribution gene C (NudC), a cochaperone of heat shock protein 90 (Hsp90), is required to stabilize filamin A in mammalian cells. Immunoprecipitation-mass spectrometry and western blotting analyses reveal that NudC interacts with filamin A. Overexpression of human NudC-L279P (an evolutionarily conserved mutation in NudC that impairs its chaperone activity) not only decreases the protein level of filamin A but also results in actin disorganization and the suppression of cell migration. Ectopic expression of filamin A is able to reverse these defects induced by the overexpression of NudC-L279P. Furthermore, Hsp90 forms a complex with filamin A. The inhibition of Hsp90 ATPase activity by either geldanamycin or radicicol decreases the protein stability of filamin A. In addition, ectopic expression of Hsp90 efficiently restores NudC-L279P overexpression-induced protein stability and functional defects of filamin A. Taken together, these data suggest NudC L279P mutation destabilizes filamin A by inhibiting the Hsp90 chaperoning pathway and suppresses cell migration.

scholarly journals TRAIP functions as an oncogene in hepatocellular carcinoma via Rb/EZH2/p21 signaling pathway

2020 ◽  
Author(s):  
Rong-Zhen Luo ◽  
Qiu-Li Li ◽  
Mei-Fang Zhang ◽  
Peng-Wei Zhang ◽  
Huimin Shen ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Biological Function ◽  
Therapeutic Potential ◽  
Ectopic Expression ◽  
Underlying Mechanism ◽  
Interacting Protein ◽  
Protein Levels ◽  
Kaplan Meier ◽  
Disease Free

Abstract Background The TRAF-interacting protein (TRAIP) has been identified as a master regulator of DNA damage and implicated in the progression of human cancers. Yet the underlying mechanism of TRAIP-mediated malignant phenotype remains unclear. Methods The expression of TRAIP in hepatocellular carcinoma (HCC) was examined by qRT-PCR, western blot and immunohistochemistry. The clinical significance of TRAIP was determined by Kaplan-Meier survival analyses. The biological function and the underlying mechanism of TRAIP in HCC progression were investigated, using cellular and molecular biological experiments.Results Here, we show that TRAIP is upregulated in HCC and functions as an oncogene via Rb/EZH2/p21 signaling. Overexpression of TRAIP, at both mRNA and protein levels, is correlated with more aggressive clinicopathological features, and unfavorable overall and disease-free survivals. In vitro experiments demonstrate that ectopic expression of TRAIP enhances, whereas knockdown of TRAIP attenuates HCC cell proliferation. Further data show that TRAIP interacts with SPAG5 in HCC cells, which results in the stabilization of TRAIP protein. TRAIP overexpression suppresses the expression of Rb, subsequently leading to the increase of EZH2 and decrease of p21. Re-expression of Rb and p21 significantly inhibits TRAIP-mediated cell growth. Conclusion Collectively, our findings suggest TRAIP exert oncogenic activity and have prognostic and therapeutic potential in HCC.

scholarly journals Mechanical stimulation promotes enthesis injury repair by mobilizing Prx1+ cells via ciliary TGF-β signaling

2021 ◽  
Author(s):  
Han Xiao ◽  
Tao Zhang ◽  
Chang Jun Li ◽  
Yong Cao ◽  
Lin Feng Wang ◽  
...  
Keyword(s):  
Cell Migration ◽  
Rotator Cuff ◽  
Single Cell ◽  
Mechanical Stimulation ◽  
Essential Role ◽  
Primary Cilia ◽  
Underlying Mechanism ◽  
Rna Sequence ◽  
Injury Repair ◽  
Tgf Β1

Proper mechanical stimulation can improve rotator cuff enthsis injury repair. However, the underlying mechanism of mechanical stimulation promoting injury repair is still unknown. In this study, we found that Prx1+ cell was essential for murine rotator cuff enthesis development identified by single-cell RNA sequence and involved in the injury repair. Proper mechanical stimulation could promote the migration of Prx1+ cells to enhance enthesis injury repair. Meantime, TGF-β signaling and primary cilia played an essential role in mediating mechanical stimulation signaling transmission. Proper mechanical stimulation enhanced the release of active TGF-β1 to promote migration of Prx1+ cells. Inhibition of TGF-β signaling eliminated the stimulatory effect of mechanical stimulation on Prx1+ cell migration and enthesis injury repair. In addition, knockdown of Pallidin to inhibit TGF-βR2 translocation to the primary cilia or deletion of IFT88 in Prx1+ cells also restrained the mechanics-induced Prx1+ cells migration. These findings suggested that mechanical stimulation could increase the release of active TGF-β1 and enhance the mobilization of Prx1+ cells to promote enthesis injury repair via ciliary TGF-β signaling.

scholarly journals Thrombomodulin is an ezrin‐interacting protein that controls epithelial morphology and promotes collective cell migration

2012 ◽  
Vol 26 (8) ◽  
pp. 3440-3452 ◽  
Author(s):  
Yun‐Yan Hsu ◽  
Guey‐Yueh Shi ◽  
Cheng‐Hsiang Kuo ◽  
Shu‐Lin Liu ◽  
Ching‐Ming Wu ◽  
...  
Keyword(s):  
Cell Migration ◽  
Collective Cell Migration ◽  
Interacting Protein ◽  
Epithelial Morphology

scholarly journals A Mammalian Ortholog of Saccharomyces cerevisiae Vac14 That Associates with and Up-Regulates PIKfyve Phosphoinositide 5-Kinase Activity

2004 ◽  
Vol 24 (23) ◽  
pp. 10437-10447 ◽  
Author(s):  
Diego Sbrissa ◽  
Ognian C. Ikonomov ◽  
Jana Strakova ◽  
Rajeswari Dondapati ◽  
Krzysztof Mlak ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Kinase Activity ◽  
Ectopic Expression ◽  
Multivesicular Body ◽  
Hek293 Cells ◽  
Lipid Kinase ◽  
Morphological Alterations ◽  
Protein Levels ◽  
Interfering Rna

ABSTRACT Multivesicular body morphology and size are controlled in part by PtdIns(3,5)P2, produced in mammalian cells by PIKfyve-directed phosphorylation of PtdIns(3)P. Here we identify human Vac14 (hVac14), an evolutionarily conserved protein, present in all eukaryotes but studied principally in yeast thus far, as a novel positive regulator of PIKfyve enzymatic activity. In mammalian cells and tissues, Vac14 is a low-abundance 82-kDa protein, but its endogenous levels could be up-regulated upon ectopic expression of hVac14. PIKfyve and hVac14 largely cofractionated, populated similar intracellular locales, and physically associated. A small-interfering RNA-directed gene-silencing approach to selectively eliminate endogenous hVac14 rendered HEK293 cells susceptible to morphological alterations similar to those observed upon expression of PIKfyve mutants deficient in PtdIns(3,5)P2 production. Largely decreased in vitro PIKfyve kinase activity and unaltered PIKfyve protein levels were detected under these conditions. Conversely, ectopic expression of hVac14 increased the intrinsic PIKfyve lipid kinase activity. Concordantly, intracellular PtdIns(3)P-to-PtdIns(3,5)P2 conversion was perturbed by hVac14 depletion and was elevated upon ectopic expression of hVac14. These data demonstrate a major role of the PIKfyve-associated hVac14 protein in activating PIKfyve and thereby regulating PtdIns(3,5)P2 synthesis and endomembrane homeostasis in mammalian cells.

scholarly journals A cytosolic heat shock protein 90 and cochaperone CDC37 complex is required for RIP3 activation during necroptosis

2015 ◽  
Vol 112 (16) ◽  
pp. 5017-5022 ◽  
Author(s):  
Dianrong Li ◽  
Tao Xu ◽  
Yang Cao ◽  
Huayi Wang ◽  
Lin Li ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Heat Shock Protein 90 ◽  
Kinase Domain ◽  
Hsp90 Inhibitor ◽  
Activation Process ◽  
Interacting Protein ◽  
Tnf Α ◽  
Chemical Inhibitors ◽  
Core Components ◽  
Necrosis Factor

Receptor-interacting protein kinase 3, RIP3, and a pseudokinase mixed lineage kinase-domain like protein, MLKL, constitute the core components of the necroptosis pathway, which causes programmed necrotic death in mammalian cells. Latent RIP3 in the cytosol is activated by several upstream signals including the related kinase RIP1, which transduces signals from the tumor necrosis factor (TNF) family of cytokines. We report here that RIP3 activation following the induction of necroptosis requires the activity of an HSP90 and CDC37 cochaperone complex. This complex physically associates with RIP3. Chemical inhibitors of HSP90 efficiently block necroptosis by preventing RIP3 activation. Cells with knocked down CDC37 were unable to respond to necroptosis stimuli. Moreover, an HSP90 inhibitor that is currently under clinical development as a cancer therapy was able to prevent systemic inflammatory response syndrome in rats treated with TNF-α. HSP90 and CDC37 cochaperone complex-mediated protein folding is thus an important part of the RIP3 activation process during necroptosis.

scholarly journals NudCL2 is an Hsp90 cochaperone to regulate sister chromatid cohesion by stabilizing cohesin subunits

2018 ◽  
Author(s):  
Yuehong Yang ◽  
Wei Wang ◽  
Min Li ◽  
Wen Zhang ◽  
Yuliang Huang ◽  
...  
Keyword(s):  
Atpase Activity ◽  
Chromosome Segregation ◽  
Sister Chromatid ◽  
Mammalian Cells ◽  
Ectopic Expression ◽  
Sister Chromatid Cohesion ◽  
Chaperone Function ◽  
Chromatid Cohesion ◽  
Protein Instability ◽  
The Stability

AbstractSister chromatid cohesion plays a key role in ensuring precise chromosome segregation during mitosis, which is mediated by the multisubunit complex cohesin. However, the molecular regulation of cohesin subunits stability remains unclear. Here, we show that NudCL2 (NudC-like protein 2) is essential for the stability of cohesin subunits by regulating Hsp90 ATPase activity in mammalian cells. Depletion of NudCL2 induces mitotic defects and premature sister chromatid separation and destabilizes cohesin subunits that interact with NudCL2. Similar defects are also observed upon inhibition of Hsp90 ATPase activity. Interestingly, ectopic expression of Hsp90 efficiently rescues the protein instability and functional deficiency of cohesin induced by NudCL2 depletion, but not vice versa. Moreover, NudCL2 not only binds to Hsp90, but also significantly modulates Hsp90 ATPase activity and promotes the chaperone function of Hsp90. Taken together, these data suggest that NudCL2 is a previously undescribed Hsp90 cochaperone to modulate sister chromatid cohesion by stabilizing cohesin subunits, providing a hitherto unrecognized mechanism that is crucial for faithful chromosome segregation during mitosis.

scholarly journals Heterogenous adhesion of follower cells affects tension profile and velocity of leader cells in primary keratocyte collective cell migration

2020 ◽  
Author(s):  
Baishali Mukherjee ◽  
Madhura Chakraborty ◽  
Arikta Biswas ◽  
Rajesh Kumble Nayak ◽  
Bidisha Sinha
Keyword(s):  
Cell Migration ◽  
Single Cell ◽  
Cell Interactions ◽  
Collective Cell Migration ◽  
Membrane Tension ◽  
High Cell ◽  
Cell Sheets ◽  
Membrane Fluctuations ◽  
High Tension ◽  
Cell To Cell Variability

AbstractSingle cell studies demonstrate membrane tension to be a central regulator of lamellipodia-driven motility bringing in front-coherence. During collective cell migration, however, tension mapping or existence of intracellular tension-gradients and the effect of cell-cell interactions have remained unexplored. In this study of membrane fluctuations and fluctuation-tension of migrating primary keratocyte cell-sheets, we first show that some leader cells are followed by followers which remain de-adhered from the substrate while being attached to other cells and thus appear to be “taking a ride”. A subtle yet significant enhanced long-timescale velocity in these leaders indicate increased directionality. Intriguingly, such leaders mostly have front-high tension gradients like single keratocytes, while followers and other leaders usually display front-low membrane tension gradients. The front-high tension gradient and higher membrane tension observed in these leaders, despite the high cell-to-cell variability in membrane tension demonstrate how leader-follower interactions and heterogenous adhesion profiles are key in collective cell migration.

scholarly journals Effect of Geometric Curvature on Collective Cell Migration in Tortuous Microchannel Devices

Micromachines ◽  
2020 ◽  
Vol 11 (7) ◽  
pp. 659
Author(s):  
Mazlee Bin Mazalan ◽  
Mohamad Anis Bin Ramlan ◽  
Jennifer Hyunjong Shin ◽  
Toshiro Ohashi
Keyword(s):  
Tissue Engineering ◽  
Cell Migration ◽  
Underlying Mechanism ◽  
Collective Cell Migration ◽  
Tissue Scaffold ◽  
Cell Behaviors ◽  
Mechanical Signals ◽  
Geometrical Constraints ◽  
Engineered Tissue ◽  
Microfabrication Technology

Collective cell migration is an essential phenomenon in many naturally occurring pathophysiological processes, as well as in tissue engineering applications. Cells in tissues and organs are known to sense chemical and mechanical signals from the microenvironment and collectively respond to these signals. For the last few decades, the effects of chemical signals such as growth factors and therapeutic agents on collective cell behaviors in the context of tissue engineering have been extensively studied, whereas those of the mechanical cues have only recently been investigated. The mechanical signals can be presented to the constituent cells in different forms, including topography, substrate stiffness, and geometrical constraint. With the recent advancement in microfabrication technology, researchers have gained the ability to manipulate the geometrical constraints by creating 3D structures to mimic the tissue microenvironment. In this study, we simulate the pore curvature as presented to the cells within 3D-engineered tissue-scaffolds by developing a device that features tortuous microchannels with geometric variations. We show that both cells at the front and rear respond to the varying radii of curvature and channel amplitude by altering the collective migratory behavior, including cell velocity, morphology, and turning angle. These findings provide insights into adaptive migration modes of collective cells to better understand the underlying mechanism of cell migration for optimization of the engineered tissue-scaffold design.

scholarly journals HAX1 impact on collective cell migration, cell adhesion, and cell shape is linked to the regulation of actomyosin contractility

2019 ◽  
Vol 30 (25) ◽  
pp. 3024-3036 ◽  
Author(s):  
Anna Balcerak ◽  
Alicja Trebinska-Stryjewska ◽  
Maciej Wakula ◽  
Mateusz Chmielarczyk ◽  
Urszula Smietanka ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Migration ◽  
Single Cell ◽  
Cancer Progression ◽  
Cancer Cell Line ◽  
Cell Junctions ◽  
Collective Cell Migration ◽  
Actomyosin Contractility ◽  
Epithelial Cell Layer ◽  
Cell Cell

HAX1 protein is involved in the regulation of apoptosis, cell motility and calcium homeostasis. Its overexpression was reported in several tumors, including breast cancer. This study demonstrates that HAX1 has an impact on collective, but not single-cell migration, thus indicating the importance of cell–cell contacts for the HAX1-mediated effect. Accordingly, it was shown that HAX1 knockdown affects cell–cell junctions, substrate adhesion, and epithelial cell layer integrity. As demonstrated here, these effects can be attributed to the modulation of actomyosin contractility through changes in RhoA and septin signaling. Additionally, it was shown that HAX1 does not influence invasive potential in the breast cancer cell line, suggesting that its role in breast cancer progression may be linked instead to collective invasion of the epithelial cells but not single-cell dissemination.

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