PREDICTION OF TORC2 PHOSPHORYLATION SITES AND ASSOCIATED PROTEINS

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。在最近的一次高通量表达筛选中，我们鉴定了一系列CREB共激活因子，称为调节CREB活性的转导因子(Torcs)。这三个TORC家族成员共享一个高度保守的N端螺旋线圈结构域，该结构域与CREB的bZIP结构域直接相关。在没有信号的情况下，TORC2以潜在的共激活因子的形式存在于细胞质中，并在cAMP和钙信号的作用下被输入细胞核。TORC2的穿梭是由相反的信号调节激酶和磷酸酶活性以及通过与14-3-3蛋白的相互作用来控制的。这项合作的目的是确定Torc2中影响其细胞质/核穿梭的结构性和受调控的磷酸化位点，并利用质谱学确定相关的激酶和磷酸酶活性。

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. In a recent high-throughput expression screen to identify novel modulators of CREB activity, we characterized a family of CREB coactivators referred to as transducers of regulated CREB activity (TORCs). The three TORC family members share a highly conserved N-terminal coiled-coil domain that mediates a direct association with the bZIP domain of CREB. In the absence of signal, TORC2 resides in the cytoplasm as a latent co-activator and is imported into the nucleus in response to cAMP and calcium signals. TORC2 shuttling is controled by opposing signal regulated kinase and phosphatase activites and through an interaction with 14-3-3 proteins. The purpose of this collaboration is to identify constitutive and regulated phosphorylation sites within Torc2 that effect its cytoplasmic/nuclear shuttling and to identify the associated kinase and phosphatase activities using mass spectrometry.