RAC P21 IS INVOLVED IN INSULIN-INDUCED MEMBRANE RUFFLING AND RHO P21 IS INVOLVED IN HEPATOCYTE GROWTH FACTOR-INDUCED AND 12-O-TETRADECANOYLPHORBOL-13-ACETATE (TPA)-INDUCED MEMBRANE RUFFLING IN KB CELLS
RAC P21 IS INVOLVED IN INSULIN-INDUCED MEMBRANE RUFFLING AND RHO P21 IS INVOLVED IN HEPATOCYTE GROWTH FACTOR-INDUCED AND 12-O-TETRADECANOYLPHORBOL-13-ACETATE (TPA)-INDUCED MEMBRANE RUFFLING IN KB CELLS
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DOI：
10.1128/mcb.14.4.2447
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发表时间：
1994-04-01
影响因子：
5.3
通讯作者：
TAKAI, Y
中科院分区：
文献类型：
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作者：
NISHIYAMA, T;SASAKI, T;TAKAI, Y
胰岛素和肝细胞生长因子（HGF）诱导KB细胞出现形态学上不同的膜皱褶。胰岛素诱导的膜皱褶被抑制rho GDI，抑制性GDP/GTP交换调节rho p21和rac p21小GTP结合蛋白的微量注射，但不抑制肉毒杆菌外切酶C3，已知选择性ADP核糖基化rho p21和损害其功能。这种rho GDI作用通过与鸟苷5 '-（3-O-硫代）三磷酸（GTP γ S）结合的rac 1 p21共同显微注射来阻止。相反，HGF诱导的膜皱褶被rho GDI或C3的微量注射抑制。通过显微注射GTP γ S结合的rhoA p21来阻止rho GDI作用，并且通过显微注射GTP γ S结合的rhoA 1 le-41 p21来阻止C3作用，rhoA 1 le-41 p21对C3具有抗性。在没有生长因子的情况下，单独显微注射GTP γ S结合的rac 1 p21或rhoA p21诱导膜皱褶。rac 1 p21诱导的膜皱褶在形态上类似于胰岛素诱导的类型，而rhoA p21诱导的皱褶明显不同于胰岛素和HGF诱导的类型。膜皱褶也诱导12-O-十四烷酰基佛波醇-13-乙酸酯（TPA），蛋白激酶C激活佛波醇酯，但不是由钙离子载体或显微注射的显性活性Ki-ras p21突变体（Ki-rasVal-12 p21）。佛波酯诱导的膜皱褶在形态上类似于rhoA p21诱导的种类，并通过微量注射rho GDI或C3抑制。这些结果表明，外消旋p21和rho GDI参与胰岛素诱导的膜皱褶，rho p21和rho GDI参与HGF和佛波酯诱导的膜皱褶。
Insulin and hepatocyte growth factor (HGF) induced morphologically different membrane rufflings in KB cells. Insulin-induced membrane ruffling was inhibited by microinjection of rho GDI, an inhibitory GDP/GTP exchange regulator for both rho p21 and rac p21 small GTP-binding proteins, but not inhibited by microinjection of botulinum exoenzyme C3, known to selectively ADP-ribosylate rho p21 and to impair its function. This rho GDI action was prevented by comicroinjection with guanosine 5'-(3-O-thio)triphosphate (GTPgammaS)-bound rac1 p21. In contrast, HGF-induced membrane ruffling was inhibited by microinjection of rho GDI or C3. This rho GDI action was prevented by comicroinjection with GTPgammaS-bound rhoA p21, and this C3 action was prevented by comicroinjection with GTPgammaS-bound rhoAIle-41 p21, which is resistant to C3. Microinjection of either GTPgammaS-bound rac1 p21 or rhoA p21 alone induced membrane ruffling in the absence of the growth factors. The rac1 p21-induced membrane ruffling was morphologically similar to the insulin-induced kind, whereas rhoA p21-induced ruffling was apparently different from both the insulin- and HGF-induced kinds. Membrane ruffling was also induced by 12-O-tetradecanoylphorbol-13-acetate (TPA), a protein kinase C-activating phorbol ester, but not by Ca2+ ionophore or microinjection of a dominant active Ki-ras p21 mutant (Ki-rasVal-12 p21). The phorbol ester-induced membrane ruffling was morphologically similar to the rhoA p21-induced kind and inhibited by microinjection of rho GDI or C3. These results indicate that rac p21 and rho GDI are involved in insulin-induced membrane ruffling and that rho p21 and rho GDI are involved in HGF- and phorbol ester-induced membrane rufflings.