Phosphatidylinositol 3-kinase is necessary and sufficient for insulin-stimulated stress fiber breakdown

被引:44
作者
Martin, SS
Rose, DW
Saltiel, AR
Klippel, A
Williams, LT
Olefsky, JM
机构
[1] UNIV CALIF SAN DIEGO, VET ADM MED CTR, DEPT MED, DIV ENDOCRINOL & METAB, LA JOLLA, CA 92093 USA
[2] WARNER LAMBERT PARKE DAVIS, PARKE DAVIS PHARMACEUT RES DIV, BIOMED SCI DOCTORAL PROGRAM, ANN ARBOR, MI 48105 USA
[3] WARNER LAMBERT PARKE DAVIS, PARKE DAVIS PHARMACEUT RES DIV, WHITTIER DIABET PROGRAM, ANN ARBOR, MI 48105 USA
[4] WARNER LAMBERT PARKE DAVIS, PARKE DAVIS PHARMACEUT RES DIV, DEPT SIGNAL TRANSDUCT, ANN ARBOR, MI 48105 USA
[5] CHIRON CORP, EMERYVILLE, CA 94608 USA
关键词
D O I
10.1210/en.137.11.5045
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Rat-1 fibroblasts overexpressing the human insulin receptor undergo rapid actin rearrangement in response to insulin. Breakdown of stress fibers present in quiescent cells is followed by transient membrane ruffling and a return of stress fibers. We investigated the signaling pathways that mediate this insulin-stimulated reorganization of the actin cytoskeleton, which was visualized with rhodamine-phalloidin. Treatment of cells with the phosphatidylinositol 3-kinase (PI3-kinase) inhibitor wortmannin prevented insulin action at the preliminary step of stress fiber breakdown. Cellular microinjection of a polyclonal antibody directed against the p85 subunit of PI3-kinase as well as a purified recombinant p85-SH2 domain protein also inhibited actin reorganization. Transient expression of a constitutively active form of PI3-kinase (p110*) was sufficient to cause both stress fiber breakdown and membrane ruffling in the absence of insulin. Microinjection of a polyclonal anti-She antibody or dominant negative N17-Ras protein did not affect actin dynamics, and although constitutively active Vla-Ras caused modest cytoskeletal reorganization, this effect was blocked by pretreatment with wortmannin. In summary, activation of PI3-kinase is necessary and sufficient to stimulate actin rearrangement, indicating that PI3-kinase may initiate the only signaling cascade required for insulin to induce cytoskeletal restructuring.
引用
收藏
页码:5045 / 5054
页数:10
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