Effects of structure of Rho GTPase-activating protein DLC-1 on cell morphology and migration.
DLC-1 encodes a Rho GTPase-activating protein (RhoGAP) and negative regulator of specific Rho family proteins (RhoA-C and Cdc42). DLC-1 is a multi-domain protein, with the RhoGAP catalytic domain flanked by an amino-terminal sterile alpha motif (SAM) and a carboxyl-terminal START domain. The roles of these domains in the regulation of DLC-1 function remain to be determined. We undertook a structure-function analysis involving truncation and missense mutants of DLC-1. We determined that the amino-terminal SAM domain functions as an autoinhibitory domain of intrinsic RhoGAP activity. Additionally, we determined that the SAM and START domains are dispensable for DLC-1 association with focal adhesions. We then characterized several mutants for their ability to regulate cell migration and identified constitutively activated and dominant negative mutants of DLC-1. We report that DLC-1 activation profoundly alters cell morphology, enhances protrusive activity, and can increase the velocity but reduce directionality of cell migration. Conversely, the expression of the amino-terminal domain of DLC-1 acts as a dominant negative and profoundly inhibits cell migration by displacing endogenous DLC-1 from focal adhesions.
Kim
Tai Young
TY
Department of Pharmacology, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
Healy
Kevin D
KD
Der
Channing J
CJ
Sciaky
Noah
N
Bang
Yung-Jue
YJ
Juliano
Rudy L
RL
D000199:Actins / Q000737:chemistry; D020816:Amino Acid Motifs; D002460:Cell Line; D002465:Cell Movement; D048430:Cell Shape; D022001:Focal Adhesions; D020690:GTPase-Activating Proteins / Q000378:metabolism; D005799:Genes, Dominant; D006801:Humans; D008954:Models, Biological; D009154:Mutation; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship; D025521:Tumor Suppressor Proteins / Q000737:chemistry* / Q000502:physiology; D020741:rho GTP-Binding Proteins / Q000378:metabolism*
D000199:Actins; C112352:DLC1 protein, human; D020690:GTPase-Activating Proteins; D025521:Tumor Suppressor Proteins; C085379:rho GTPase-activating protein; D020741:rho GTP-Binding Proteins
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