Ras‐interacting domain of RGL blocks Ras‐dependent signal transduction in Xenopus oocytes

FEBS Letters

Koyama, Shinya²  Williams, Lewis T.¹  Chen, Yen-Wen¹  Kikuchi, Akira²  Muslin, Anthony J.¹  Ikeda, Masahiro² 

[1] Cardiovascular Research Institute and Daiichi Research Center, University of California, San Francisco, CA 94143-0130, USA;Department of Biochemistry, Hiroshima University School of Medicine, 1-2-3 Kasumi, Minami-ku, Hiroshima 734, Japan

关键词: RID;    RGL;    Ras;    Extracellular signal-regulated kinase;    Xenopus oocyte;    GAP;    GTPase activating protein;    PI3-kinase;    phosphatidylinositol 3-kinase;    MEKK;    MEK kinase;    ERK;    extracellular signal-regulated kinase;    RGL;    RalGDS like;    RID;    Ras-interacting domain;    NF-1;    neurofibromatosis 1;    HA;    hemaggulutinin;    PCR;    polymerase chain reaction;    GST;    glutathione-S-transferase;    Sf9;    Spodoptera frugiperda;    E. coli;    Escherichia coli;    DTT;    dithiothreitol;    BSA;    bovine serum albumin;    GVBD;    germinal vesicle breakdown;    MEK;    mitogen-activated protein kinase and ERK kinase;    SAPK;    stress-activated protein kinase;   

DOI  :  10.1016/0014-5793(96)00018-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

RalGDS family members (RalGDS and RGL) interact with the GTP-bound form of Ras through its effector loop. The C-terminal region (amino acids 602–768) of RGL is responsible for binding to Ras. In this paper we characterized a Ras-interacting domain of RGL using deletion mutants of RGL(602–768). RGL(602–768), RGL(632–768), and RGL(602–734) bound to the GTP-bound form of Ras and inhibited the GAP activity of NF-1. RGL(646–768) showed a low binding activity to Ras and inhibited GAP activity of NF-1 weakly. None of RGL(659–768), RGL(685–768), RGL(602–709), and RGL(602–686) bound to Ras or inhibited GAP activity of NF-1. These results indicate that amino acids 632–734 of RGL constitute a nearly minimal domain that contains the binding element for Ras. RGL(632–734) inhibited v-Ras- but not progesterone-induced Xenopus oocyte maturation. Furthermore, RGL(632–734) inhibited v-Ras- but not v-Raf-dependent extracellular signal-regulated kinase activation in Xenopus oocytes. These results clearly demonstrate that the Ras-interacting domain of RGL is important for Ras-dependent signal transduction in vivo.

【 授权许可】

Unknown