IDENTIFICATION OF A MOUSE P21(CDC42/RAC) ACTIVATED KINASE
IDENTIFICATION OF A MOUSE P21(CDC42/RAC) ACTIVATED KINASE
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DOI：
10.1074/jbc.270.39.22731
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发表时间：
1995-09-29
影响因子：
4.8
通讯作者：
CERIONE, RA
中科院分区：
文献类型：
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作者：
BAGRODIA, S;TAYLOR, SJ;CERIONE, RA
我们从小鼠成纤维细胞 cDNA 文库中分离出哺乳动物 PAI（p21 激活激酶）和酵母 Ste20 丝氨酸/苏氨酸激酶家族的新成员，命名为 mPAK-3。在酿酒酵母中表达 mPAK-3 可部分恢复 ste20 无效细胞中的交配功能。与其他 PAK 一样，mPAK-3 包含假定的 Cdc42Hs/Rac 结合序列，并且在 COS 中瞬时表达时在细胞中，全长 mPAK-3 结合激活的（GTP gamma S（鸟苷 5'-3-O-（硫代三磷酸）结合）谷胱甘肽 S-转移酶 (GST)-Cdc42Hs 和 GST-Rac1，但不结合 GST-RhoA。正如对推定靶分子所预期的那样，mPAX-3 不结合 Cdc42Hs 的效应结构域突变体。此外，激活的 His 标记的 Cdc42Hs有趣的是，mPAK-3 的氨基末端区域含有潜在的 SH3 结合位点，并且我们发现体外和体内表达的 mPAK-3 显示出与磷脂酶 C-gamma 的 SH3 结构域和接头蛋白 Nck 中的至少一个 SH3 结构域的高度特异性结合。体内PAK家族的成员。
We have isolated a novel member of the mammalian PAI( (p21 activated kinase) and yeast Ste20 serine/threonine kinase family from a mouse fibroblast cDNA library, designated mPAK-3. Expression of mPAK-3 in Saccharomyces cerevisiae partially restores mating function in ste20 null cells. Like other PAKs, mPAK-3 contains a putative Cdc42Hs/Rac binding sequence and when transiently expressed in COS cells, full-length mPAK-3 binds activated (GTP gamma S (guanosine 5'-3-O-(thiotriphosphate)-bound) glutathione S-transferase (GST)-Cdc42Hs and GST-Rac1 but not GST-RhoA. As expected for a putative target molecule, mPAX-3 does not bind to an effector domain mutant of Cdc42Hs. Furthermore, activated His-tagged Cdc42Hs and His-tagged Rac stimulate mPAK-3 autophosphorylation and phosphorylation of myelin basic protein by mPAK-3 in vitro. Interestingly, the amino-terminal region of mPAK-3 contains potential SH3-binding sites and we find that mPAK-3, expressed in vitro and in vivo, shows highly specific binding to the SH3 domain of phospholipase C-gamma and at least one SH3 domain in the adapter protein Nck. These results raise the possibility of an additional level of regulation of the PAK family in vivo.