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包含磷脂酰肌醇特异性磷脂酶C-γ和Src的SH2/SH3区域的v-Crk嵌合体的生物学和生化活性

Biological and biochemical activity of v-Crk chimeras containing the SH2/SH3 regions of phosphatidylinositol-specific phospholipase C-gamma and Src.

作者信息

Matsuda M, Reichman C T, Hanafusa H

机构信息

Rockefeller University, New York, New York 10021-6399.

出版信息

J Virol. 1992 Jan;66(1):115-21. doi: 10.1128/JVI.66.1.115-121.1992.

DOI:10.1128/JVI.66.1.115-121.1992
PMID:1370084
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC238266/
Abstract

The chicken CT10 virus oncogene product, P47gag-crk, contains SH2/SH3 domains that have been identified as conserved domains among proteins involved in signal transduction. We studied the functional similarity of the SH2/SH3 domains by replacing those of v-Crk with those of phosphatidylinositol-specific phospholipase C-gamma, v-Src, or c-Src. The transforming activity of v-Crk was partially retained in a mutant with a v-Src SH3 domain but not in the other mutants with heterologous SH2/SH3 domains. Mutant viruses with Crk-SH2/SH2' domains induced tyrosine phosphorylation of cellular proteins, but mutants with phosphatidylinositol-specific phospholipase C-gamma or Src SH2/SH2' domains did not. However, the mutant proteins with heterologous SH2/SH2' regions were able to weakly associate with some phosphotyrosine-containing proteins in vitro. These results indicate that in the context of the P47gag-crk structure, the requirement of Crk-SH2/SH3 is more stringent for its activity to induce cell transformation than to cause phosphorylation of cellular proteins. The substitution with heterologous sequences least perturbs the capacity to bind phosphotyrosine-containing proteins. In each case, the SH3 domain is more flexible to substitution than is the SH2 domain.

摘要

鸡CT10病毒癌基因产物P47gag-crk含有SH2/SH3结构域，这些结构域已被确定为参与信号转导的蛋白质中的保守结构域。我们通过用磷脂酰肌醇特异性磷脂酶C-γ、v-Src或c-Src的SH2/SH3结构域替换v-Crk的相应结构域，研究了SH2/SH3结构域的功能相似性。v-Crk的转化活性在具有v-Src SH3结构域的突变体中部分保留，但在具有异源SH2/SH3结构域的其他突变体中则没有。具有Crk-SH2/SH2'结构域的突变病毒诱导细胞蛋白的酪氨酸磷酸化，但具有磷脂酰肌醇特异性磷脂酶C-γ或Src SH2/SH2'结构域的突变体则不能。然而，具有异源SH2/SH2'区域的突变蛋白在体外能够与一些含磷酸酪氨酸的蛋白弱结合。这些结果表明，在P47gag-crk结构的背景下，Crk-SH2/SH3对其诱导细胞转化活性的要求比对引起细胞蛋白磷酸化的要求更为严格。用异源序列替换对结合含磷酸酪氨酸蛋白的能力干扰最小。在每种情况下，SH3结构域比SH2结构域对替换更具灵活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06b4/238266/78e75cca75af/jvirol00034-0139-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06b4/238266/8d8cedab3131/jvirol00034-0137-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06b4/238266/2300d761ee4a/jvirol00034-0138-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06b4/238266/8f13c877ccc5/jvirol00034-0138-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06b4/238266/8e0c608dd409/jvirol00034-0138-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06b4/238266/73ac44acaeb7/jvirol00034-0139-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06b4/238266/78e75cca75af/jvirol00034-0139-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06b4/238266/8d8cedab3131/jvirol00034-0137-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06b4/238266/2300d761ee4a/jvirol00034-0138-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06b4/238266/8f13c877ccc5/jvirol00034-0138-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06b4/238266/8e0c608dd409/jvirol00034-0138-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06b4/238266/73ac44acaeb7/jvirol00034-0139-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06b4/238266/78e75cca75af/jvirol00034-0139-b.jpg

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