Regulation of integrin expression by Gα12: An additional potential mechanism modulating cell attachment.
Integrins regulate cell attachment and migration through interactions with specific proteins in the extra-cellular matrix. Heterotrimeric G proteins are essential signal transduction proteins that intersect with integrin signaling to regulate fundamental cellular behaviors. Although integrin and G protein signaling often act in concert, how these mechanisms interact in epithelial cells has not been extensively studied. We recently reported Gα12 regulation of epithelial cell attachment and migration on collagen-I through α2β1 integrins (Kong et al. Mol Biol Cell 2009). Activated Gα12 inhibited α2β1 integrin functions through an inside-out signaling mechanism that involved Rho, Src and protein phosphatases without affecting α2 or β1 expression. Activated Gα12 prevented tubulogenesis in 3D-MDCK cell cultures and promoted the formation of cystic structures. Herein, we extend these findings to show Gα12-stimulated transcriptional changes in integrin expression that affect MDCK cell attachment. Based on results from a microarray with MDCK cells expressing constitutively active Gα12 (QLα12), we confirmed with real time PCR that expressing QLα12 led to a 4-fold inhibition of α6 mRNA expression. Cell surface expression and total α6 protein was reduced by FACS and immunofluorescence. QLα12 expressing MDCK cells also revealed less attachment to laminin-5, an α6 integrin ligand. Taken together, G proteins regulate integrins through canonical signaling pathways and potentially regulate integrin expression levels to modulate cellular responses in a variety of pathophysiologic conditions including polycystic kidney disease.
Kong
Tianqing
T
Renal Division, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA. tkong@rics.bwh.harvard.edu
Xu
Daosong
D
Tran
Mei
M
Denker
Bradley M
BM
D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D004285:Dogs; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism*; D044365:GTP-Binding Protein alpha Subunits, G12-G13 / Q000378:metabolism*; D006801:Humans; D016023:Integrins / Q000378:metabolism*; D016891:Polycystic Kidney, Autosomal Dominant / Q000378:metabolism / Q000473:pathology; D015398:Signal Transduction
D016023:Integrins; D044365:GTP-Binding Protein alpha Subunits, G12-G13