The headpiece domain of dematin regulates cell shape, motility, and wound healing by modulating RhoA activation.
RhoA is known to participate in cytoskeletal remodeling events through several signaling pathways, yet the precise mechanism of its activation remains unknown. Here, we provide the first evidence that dematin functions upstream of RhoA and regulates its activation. Primary mouse embryonic fibroblasts were generated from a dematin headpiece domain null (HPKO) mouse, and the visualization of the actin morphology revealed a time-dependent defect in stress fiber formation, membrane protrusions, cell motility, and cell adhesion. Rescue experiments using RNA interference and transfection assays revealed that the observed phenotypes are due to a null effect and not a gain of function in the mutant fibroblasts. In vivo wounding of adult HPKO mouse skin showed a decrease in wound healing (reepithelialization and granulation) compared to the wild-type control. Biochemical analysis of the HPKO fibroblasts revealed a sustained hyperphosphorylation of focal adhesion kinase (FAK) at tyrosine 397 as well as a twofold increase in RhoA activation. Inhibition of both RhoA and FAK signaling using C3 toxin and FRNK (focal adhesion kinase nonrelated kinase), respectively, revealed that dematin acts upstream of RhoA. Together, these results unveil a new function of dematin as a negative regulator of the RhoA activation pathway with physiological implications for normal and pathogenic signaling pathways.
Mohseni
Morvarid
M
Department of Pharmacology, UIC Cancer Center, University of Illinois College of Medicine, Chicago, IL 60612-3725, USA.
Chishti
Athar H
AH
D000818:Animals; D001798:Blood Proteins / Q000737:chemistry* / Q000378:metabolism*; D002448:Cell Adhesion; D002462:Cell Membrane / Q000201:enzymology; D002465:Cell Movement; D048430:Cell Shape; D003598:Cytoskeletal Proteins; D004789:Enzyme Activation; D005347:Fibroblasts / Q000166:cytology* / Q000201:enzymology*; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D010750:Phosphoproteins / Q000737:chemistry* / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D034741:RNA, Small Interfering / Q000378:metabolism; D014162:Transfection; D014945:Wound Healing; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
D001798:Blood Proteins; D003598:Cytoskeletal Proteins; C489041:Dmtn protein, mouse; D010750:Phosphoproteins; D034741:RNA, Small Interfering; D051416:Focal Adhesion Protein-Tyrosine Kinases; D020742:rhoA GTP-Binding Protein