BCCIP associates with the receptor protein tyrosine phosphatase PTPmu.
BCCIP associates with the receptor protein tyrosine phosphatase PTPmu.
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DOI：
10.1002/jcb.21907
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发表时间：
2008-11-01
影响因子：
4
通讯作者：
Brady-Kalnay, Susann M.
中科院分区：
文献类型：
--
作者：
Phillips-Mason, Polly J.;Mourton, Tracy;Major, Denice L.;Brady-Kalnay, Susann M.
受体蛋白酪氨酸磷酸酶PTPmu属于粘附分子家族，其在其细胞外片段中含有细胞-细胞粘附基序，在其细胞内片段中含有催化结构域。PTPmu介导粘附和表现出酶活性的能力使得PTPmu成为响应细胞-细胞粘附的信号转导的优秀候选者。为了鉴定PTPmu的下游信号传导伙伴，我们使用PTPmu的第一个酪氨酸磷酸酶结构域作为诱饵进行了改良的酵母双杂交筛选。我们从HeLa细胞文库中分离了一个编码BRCA 2和CDKN 1A（CDKN 1A [p21]）相互作用蛋白（BCCIP）的相互作用克隆。BCCIP是一种p21和BRCA 2相互作用蛋白，已被证明在细胞周期阻滞和DNA修复中发挥作用。在这篇手稿中，我们确认BCCIP和PTPmu之间的相互作用，在酵母中使用体外生化研究和表征BCCIP作为PTPmu结合蛋白。我们证明，BCCIP是由Src酪氨酸激酶磷酸化和去磷酸化的PTPmu酪氨酸磷酸酶在体外。此外，我们表明，BCCIP是必需的PTPmu的神经突起生长试验中的允许和排斥功能，表明BCCIP和PTPmu是在一个共同的信号转导途径。
The receptor protein tyrosine phosphatase PTPmu belongs to a family of adhesion molecules that contain cell-cell adhesion motifs in their extracellular segments and catalytic domains within their intracellular segments. The ability of PTPmu both to mediate adhesion and exhibit enzymatic activity makes PTPmu an excellent candidate to transduce signals in response to cell-cell adhesion. In an effort to identify downstream signaling partners of PTPmu, we performed a modified yeast two-hybrid screen using the first tyrosine phosphatase domain of PTPmu as bait. We isolated an interacting clone encoding BRCA2 and CDKN1A (CDKN1A [p21]) interacting protein (BCCIP) from a HeLa cell library. BCCIP is a p21 and BRCA2 interacting protein that has been shown to play roles in both cell cycle arrest and DNA repair. In this manuscript, we confirm the interaction between BCCIP and PTPmu identified in yeast using in vitro biochemical studies and characterize BCCIP as a PTPmu binding protein. We demonstrate that BCCIP is phosphorylated by the Src tyrosine kinase and dephosphorylated by the PTPmu tyrosine phosphatase in vitro. Furthermore, we show that BCCIP is required for both the permissive and repulsive functions of PTPmu in neurite outgrowth assays, suggesting BCCIP and PTPmu are in a common signal transduction pathway.
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