Interaction of Phospholipase C-γ<sub>1</sub>with Villin Regulates Epithelial Cell Migration

Tomar, Alok; George, Sudeep P.; Kansal, Pallavi; Wang, Yaohong; Khurana, Seema

doi:10.1074/jbc.m604323200

Cited by 29 publications

(78 citation statements)

References 48 publications

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“…18 We have previously identified 10 phosphorylation sites in villin and demonstrated that mutation of these sites inhibits the actin severing activity of villin. 19 Further, we have demonstrated the absolute requirement of c-Src kinase for tyrosine phosphorylation of villin. 20 Gelsolin is also tyrosinephosphorylated by c-Src kinase, although the tyrosinephosphorylated residues and the significance of phosphorylation for the actin-regulatory functions of gelsolin have not been identified.…”

Section: Villin Inhibits Apoptosis By Preserving Actin Dynamicsmentioning

confidence: 73%

Actin reorganization as the molecular basis for the regulation of apoptosis in gastrointestinal epithelial cells

et al. 2012

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The gastrointestinal (GI) epithelium is a rapidly renewing tissue in which apoptosis represents part of the overall homeostatic process. Regulation of apoptosis in the GI epithelium is complex with a precise relationship between cell position and apoptosis. Apoptosis occurs spontaneously and in response to radiation and cytotoxic drugs at the base of the crypts. By contrast, the villus epithelial cells are extremely resistant to apoptosis. The molecular mechanism underlying this loss of function of villus epithelial cells to undergo apoptosis shortly after their exit from the crypt is unknown. In this study we demonstrate for the first time, that deletion of two homologous actin-binding proteins, villin and gelsolin renders villus epithelial cells extremely sensitive to apoptosis. Ultrastructural analysis of the villin-gelsolin À/À double-knockout mice shows an abnormal accumulation of damaged mitochondria demonstrating that villin and gelsolin function on an early step in the apoptotic signaling at the level of the mitochondria. A characterization of functional and ligand-binding mutants demonstrate that regulated changes in actin dynamics determined by the actin severing activities of villin and gelsolin are required to maintain cellular homeostasis. Our study provides a molecular basis for the regulation of apoptosis in the GI epithelium and identifies cell biological mechanisms that couple changes in actin dynamics to apoptotic cell death.

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Section: Villin Inhibits Apoptosis By Preserving Actin Dynamicsmentioning

confidence: 73%

Actin reorganization as the molecular basis for the regulation of apoptosis in gastrointestinal epithelial cells

et al. 2012

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“…MDCK or HeLa Tet-Off cells were stably transfected with HA-tagged wild-type villin protein as described previously (16). To repress the expression of villin gene, cells were cultured in the presence of 10 ng/ml doxycycline.…”

Section: Methodsmentioning

confidence: 99%

“…To repress the expression of villin gene, cells were cultured in the presence of 10 ng/ml doxycycline. We have previously demonstrated the induction of villin following doxycycline withdrawal and shown the absence of villin in the MDCK and HeLa Tet-Off cells in the presence of doxycycline (16,17). In addition, IEC-6 cells were infected with recombinant adenovirus expressing full-length human villin protein as described previously (19).…”

Section: Methodsmentioning

confidence: 99%

“…Apoptosis Assay-An enzyme-linked immunosorbent assay was used to quantitate DNA fragmentation in MDCK cells as described before (16). Cells were cultured in 12-well plates, lysed, and centrifuged to isolate nuclei.…”

Section: Methodsmentioning

confidence: 99%

“…Doxycycline treatment was used to suppress villin expression but had no effect on cell apoptosis (supplemental Fig. 1B) (16). Cells were treated with camptothecin (20 M) and monitored 0 -14 h post-treatment.…”

Section: Villin-null Mice Demonstrate Increased Epithelial Cell Apoptmentioning

confidence: 99%

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A Novel Role for Villin in Intestinal Epithelial Cell Survival and Homeostasis

Wang

Srinivasan

Siddiqui

et al. 2008

Journal of Biological Chemistry

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Apoptosis is a key regulator for the normal turnover of the intestinal mucosa, and abnormalities associated with this function have been linked to inflammatory bowel disease and colorectal cancer. Despite this, little is known about the mechanism(s) mediating intestinal epithelial cell apoptosis. Villin is an actin regulatory protein that is expressed in every cell of the intestinal epithelium as well as in exocrine glands associated with the gastrointestinal tract. In this study we demonstrate for the first time that villin is an epithelial cell-specific anti-apoptotic protein. Absence of villin predisposes mice to dextran sodium sulfate-induced colitis by promoting apoptosis. To better understand the cellular and molecular mechanisms of the anti-apoptotic function of villin, we overexpressed villin in the Madin-Darby canine kidney Tet-Off epithelial cell line to demonstrate that expression of villin protects cells from apoptosis by maintaining mitochondrial integrity thus inhibiting the activation of caspase-9 and caspase-3. Furthermore, we report that the anti-apoptotic response of villin depends on activation of the pro-survival proteins, phosphatidylinositol 3-kinase and phosphorylated Akt. The results of our studies shed new light on the previously unrecognized function of villin in the regulation of apoptosis in the gastrointestinal epithelium.

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Site‐specific phosphorylation of villin remodels the actin cytoskeleton to regulate Sendai viral glycoprotein‐mediated membrane fusion

et al. 2019

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Connivance of cellular factors during virus‐host cell membrane fusion is poorly understood. We have recently shown that cellular villin plays an important role during membrane fusion of reconstituted Sendai virosomes with hepatocytes. Here, we employed villin‐null Chinese Hamster Ovary (CHO) cells, where villin expression led to an increased fusion with virosomes, which was further enhanced due to tyrosine phosphorylation in the presence of c‐src. However, the villin RRI mutant, lacking actin‐severing function, failed to augment membrane fusion. Furthermore, quantitative mass spectrometry and detailed analysis revealed Tyr499 to be the key phosphorylation site of villin responsible for the enhancement of virosome‐CHO cell fusion. Overall, our results demonstrate a critical role for villin and its cell‐type dependent phosphorylation in regulating membrane fusion.

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Interaction of Phospholipase C-γ₁with Villin Regulates Epithelial Cell Migration

Cited by 29 publications

References 48 publications

Actin reorganization as the molecular basis for the regulation of apoptosis in gastrointestinal epithelial cells

Actin reorganization as the molecular basis for the regulation of apoptosis in gastrointestinal epithelial cells

A Novel Role for Villin in Intestinal Epithelial Cell Survival and Homeostasis

Site‐specific phosphorylation of villin remodels the actin cytoskeleton to regulate Sendai viral glycoprotein‐mediated membrane fusion

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Interaction of Phospholipase C-γ1with Villin Regulates Epithelial Cell Migration

Cited by 29 publications

References 48 publications

Actin reorganization as the molecular basis for the regulation of apoptosis in gastrointestinal epithelial cells

Actin reorganization as the molecular basis for the regulation of apoptosis in gastrointestinal epithelial cells

A Novel Role for Villin in Intestinal Epithelial Cell Survival and Homeostasis

Site‐specific phosphorylation of villin remodels the actin cytoskeleton to regulate Sendai viral glycoprotein‐mediated membrane fusion

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Product

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Interaction of Phospholipase C-γ₁with Villin Regulates Epithelial Cell Migration