PREDICTION OF TORC2 PHOSPHORYLATION SITES AND ASSOCIATED PROTEINS

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。在最近的高通量表达筛选，以确定新的CREB活性调节剂，我们的特点是CREB辅激活因子家族被称为调节CREB活性（TORC）的换能器。三个TORC家族成员共享高度保守的N-末端卷曲螺旋结构域，其介导与CREB的bZIP结构域的直接关联。在没有信号的情况下，TORC 2作为潜在的共激活剂存在于细胞质中，并响应cAMP和钙信号而被输入到细胞核中。TORC 2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Torc 2内的组成和调节磷酸化位点，影响其细胞质/核穿梭，并使用质谱法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.