POST-TRANSLATIONAL MODIFICATION OF SPD-2/5 AND SAS5/6

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 中心体是动物细胞中主要的微管组织中心， 在细胞组织、极性和分裂中起作用。中心体数目、大小 在几乎所有的人类肿瘤类型中都观察到了肿瘤的形态学变化。 四种蛋白质（SPD-2、SPD-5、PLK-1和Aurora-A激酶（AIR-1））的消耗 1))被证明有严重的缺陷，在招聘的pericentriolar材料（PCM）。 这种募集缺陷导致中心体不能使适当的细胞核化， 数量的微管，并最终经历灾难性的有丝分裂缺陷作为适当的 不形成有丝分裂纺锤体。 我们的目标是试图了解SPD-2，SPD-5，PLK-1和 通过评估AIR-1和PLK-1激酶活性对中心体的重要性， 组装，并通过使用体外和体内方法的组合来确定SPD- 2和/或SPD-5是Aurora-A激酶的激活剂或底物。 通过与生理蛋白质组学中心（CPP）的合作，我们将使用 用两种激酶AIR-1对SPD-5和SPD-2进行体外激酶测定后的质谱分析 和PLK-1来鉴定被磷酸化修饰的位点。 因此，CPP的贡献对于理解中心体组装至关重要 因此，它将影响我们对癌症、转移和 发展

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. Centrosomes, the major microtubule organizing centers in animal cells, perform critical functions in cell organization, polarity and division. Abnormalities in centrosome number, size and morphology have been observed in nearly all human tumor types. Depletion of four proteins (SPD-2, SPD-5, PLK-1 and Aurora-A kinase (AIR- 1)) was shown to have severe defects in the recruitment of pericentriolar material (PCM). This recruitment defect results in centrosomes that are unable to nucleate the proper number of microtubules and ultimately experience catastrophic mitotic defects as proper mitotic spindles do not form. Our goal is try to understand the mechanistic relationship between SPD-2, SPD-5, PLK-1 and AIR-1 by assessing the importance of the kinase activity of AIR-1 and PLK-1 for centrosome assembly, and by using a combination of in vitro and in vivo approaches to determine if SPD- 2 and/or SPD-5 are activators or susbstrates of Aurora-A kinase. Through our collaboration with The Center for Physiological Proteomics (CPP), we will use mass spectrometry after in vitro kinase assays of SPD-5 and SPD-2 with both kinases AIR-1 and PLK-1 to identify the sites that are modified by phosphorylation. The contribution from CPP will therefore be crucial to understanding centrosome assembly and function will therefore impact on our understanding of cancer, metastasis and development.