EFFECT OF PHOSPHORYLATION ON INTERACTION OF P130CAS WITH AND-34

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 一个受资助的项目（CA114094-01：人类乳腺癌中 AND-34 的分析）试图确定为什么特定蛋白质 AND-34 的过度表达会使通常依赖雌激素的人类乳腺癌细胞系对抗雌激素的生长抑制产生抵抗力。 AND-34 在小鼠 B 细胞中表达，在淋巴粘附和迁移中发挥作用。我们已经证明 AND-34 与粘着斑接头蛋白 p130Cas 结合。 AND-34 的过表达（而非其他两个密切相关的家族成员（NSP-1 和 NSP-2））会导致细胞周期蛋白 D1 启动子的激活和抗雌激素抵抗。所有三个家族成员均与 p130Cas 结合。 有一些证据表明 AND-34 的过度表达可能会影响 p130Cas 的磷酸化。尽管 p130Cas 是众所周知的 Src 家族激酶底物，但这似乎并不是通过 Src 家族激酶依赖性机制发生的。我们正在使用质谱法来确定 AND-34 过度表达后 p130Cas 磷酸化的位置。我们将比较此 AND-34 过表达细胞系中 p130Cas 的磷酸化模式与 MCF-7 细胞中观察到的磷酸化模式。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. A funded project (CA114094-01: Analysis of AND-34 in human breast cancer) seeks to establish why over-expression of a specific protein, AND-34, allows normally estrogen-dependent human breast cancer cell lines to become resistant to growth inhibition by anti-estrogens. AND-34 is expressed in murine B cells and plays a role in lymphoid adherence and migration. We have shown that AND-34 binds to a focal adhesion adapter protein, p130Cas. Over-expression of AND-34, but not two other closely related family members (NSP-1 and NSP-2) leads to activation of the cyclin D1 promoter and anti-estrogen resistance. All three family members bind to p130Cas. There is some evidence that over-expression of AND-34 may affect phosphorylation of p130Cas. This does not appear to occur by a Src family kinase-dependent mechanism, although p130Cas is a well known Src family kinase substrate. We are using mass spectrometry to establish where p130Cas is phosphorylated following over-expression of AND-34. We will compare the phosphorylation pattern of p130Cas from this AND-34 over-expressing cell line with that observed in MCF-7 cells.