Involvement of rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI) in cell motility.
Evidence is accumulating that rho p21, a ras p21-related small GTP-binding protein (G protein), regulates the actomyosin system. The actomyosin system is known to be essential for cell motility. In the present study, we examined the action of rho p21, its inhibitory GDP/GTP exchange protein (named rho GDI), its stimulatory GDP/GTP exchange protein (named smg GDS), and Clostridium botulinum ADP-ribosyltransferase C3, known to selectively ADP-ribosylate rho p21 and to impair its function, in cell motility (chemokinesis) of Swiss 3T3 cells. We quantitated the capacity of cell motility by measuring cell tracks by phagokinesis. Microinjection of the GTP gamma S-bound active form of rhoA p21 or smg GDS into Swiss 3T3 cells did not affect cell motility, but microinjection of rho GDI into the cells did inhibit cell motility. This rho GDI action was prevented by comicroinjection of rho GDI with the GTP gamma S-bound form of rhoA p21 but not with the same form of rhoA p21 lacking the C-terminal three amino acids which was not posttranslationally modified with lipids. The rho GDI action was not prevented by Ki-rasVal-12 p21 or any of the GTP gamma S-bound form of other small GTP-binding proteins including rac1 p21, G25K, and smg p21B. Among these small G proteins, rhoA p21, rac1 p21, and G25K are known to be substrates for rho GDI. The rho GDI action was not prevented by comicroinjection of rho GDI with smg GDS. Microinjection of C3 into Swiss 3T3 cells also inhibited cell motility. These results indicate that the rho GDI-rho p21 system regulates cell motility, presumably through the actomyosin system.
Takaishi
K
K
Department of Biochemistry, Kobe University School of Medicine, Japan.
Kikuchi
A
A
Kuroda
S
S
Kotani
K
K
Sasaki
T
T
Takai
Y
Y
D016475:3T3 Cells; D019883:Alkyl and Aryl Transferases; D000818:Animals; D002465:Cell Movement; D019204:GTP-Binding Proteins / Q000378:metabolism*; D020730:Guanine Nucleotide Dissociation Inhibitors; D006150:Guanine Nucleotides / Q000378:metabolism; D066298:In Vitro Techniques; D051379:Mice; D008845:Microinjections; D012323:RNA Processing, Post-Transcriptional; D014166:Transferases / Q000378:metabolism; D062947:rho-Specific Guanine Nucleotide Dissociation Inhibitors; D020742:rhoA GTP-Binding Protein
D020730:Guanine Nucleotide Dissociation Inhibitors; D006150:Guanine Nucleotides; D062947:rho-Specific Guanine Nucleotide Dissociation Inhibitors; D014166:Transferases; D019883:Alkyl and Aryl Transferases; C068462:rhoA p21 geranylgeranyltransferase; D019204:GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein