Interaction of RSK2 with 5-HT2A receptor

描述(由申请人提供)：拟议研究的目标是验证RSK2与酵母-2杂交筛选中鉴定的5-HT2A受体的第三个细胞内环的相互作用。一旦建立了这种相互作用，它的功能意义将被确定。我们的中心假设是RSK2与5-HT2A受体相互作用，表明在信号转导中发挥作用。这种相互作用很可能在5-HT2a受体对MAP-Kinase级联通路的配体依赖激活中起作用。RSK2基因的失活突变是人类Coffin-Lowry综合征的原因。这种综合征的特征是严重的非特异性精神发育迟滞、精神病和进行性骨骼畸形。Coffin-Lowry综合征中出现的精神病可能被证明归因于5-HT2A信号转导的功能障碍。为了验证这一中心思想，我们建议通过免疫共沉淀研究和共聚焦显微镜研究来验证5-HT2A受体的第三个细胞内环与RSK2的相互作用。RSK2与5-HT2A受体的结合部位将通过结合双杂交分析的定点突变研究来确定。RSK2和5-HT2A受体之间的相互作用的功能作用将被定义。

DESCRIPTION (provided by applicant): The goal of the proposed research is to verify the interaction of RSK2 with the third intracellular loop of the 5-HT2A receptor that was identified in a yeast-2-hybrid screen. Once this interaction has been established, its functional significance will be determined. Our central hypothesis is that RSK2 interacts with the 5-HT2A receptor, indicating a role in signal transduction. It is likely that this interaction plays a role in the ligand-dependent activation of the MAP Kinase cascade by the 5-HT2A receptor. Inactivating mutations in the RSK2 gene are responsible for the human Coffin-Lowry syndrome. This syndrome is characterized by severe non-specific mental retardation, psychosis, and progressive skeletal deformations. The psychosis that is present in Coffin-Lowry syndrome may prove to be attributed to a disfunction of 5-HT2A signaling. To test this central idea we propose to verify the interaction of the 5-HT2A receptor's third intracellular loop and RSK2 via coimmunoprecipitation studies and through confocal microscopy studies. The binding site of RSK2 to the 5-HT2A receptor will be defined via site-directed mutageneis studies in conjunction with 2-hybrid analysis. The functional role of the interaction between RSK2 and the 5-HT2A receptor will be defined.