Drosophila as a Model to Investigate Rett Pathogenesis

描述(由申请人提供)：编码甲基CpG结合蛋白2(MeCP2)的基因突变会导致Rett综合征(RTT)和其他一系列男性和女性的精神发育迟滞综合征。RTT是由MECP2的功能缺失突变引起的，MECP2与靶DNA结合，并招募Sin3A和组蛋白脱乙酰酶来沉默转录。MeCP2是一个更大的基因家族的一部分，即甲基CpG结合结构域(MBD)家族，该家族在苍蝇通过人类时是保守的。利用UAS-GAL4系统在果蝇中过表达MECP2会产生表型，可能是通过与MBD基因竞争来实现的。我建议继续对全长品系进行鉴定，并创造更多的转基因果蝇：两个品系过度表达在RETT患者中常见的MECP2突变，另一个品系产生不含MBD区域的MeCP2。其次，我将描述果蝇MBD基因CG10042的天然RNA和蛋白质表达模式。然后，我将通过P元件突变和与基因组Edna杂交的RNAi来研究它的功能。我将鉴定所有产生的表型，并使用这些菌株通过F1修饰物筛选来鉴定MeCP2和/或CG10042的修饰物，以深入了解MeCP2功能的途径，到目前为止，这一功能仅在体外进行研究。通过拟议的遗传学研究，我希望找出可以研究的蛋白质，以研究RETT及相关疾病的精神和神经功能障碍的机制。

DESCRIPTION (provided by applicant): Mutations in the gene encoding methyl-CpG-binding protein 2 (MeCP2) cause Rett syndrome (RTT) and a host of other mental retardation syndromes in both males and females. RTT results from loss-of-function mutations of MECP2, which binds to target DNA and recruits Sin3A and histone deacetylases to silence transcription. MeCP2 is part of a larger family of genes, the methyl-CpG-binding domain (MBD) family, which is conserved from fly through humans. Overexpression of MECP2 in Drosophila using the UAS-GAL4 system produces a phenotype, possibly by competing with MBD genes. I propose to continue characterization of the full-length lines and create additional transgenic flies: two lines which overexpress MECP2 mutations commonly found in Rett patients and one line which produces MeCP2 without the MBD region. Secondly, I will characterize the Drosophila MBD gene CG10042 for native RNA and protein expression patterns. Then I will investigate its function through P-element mutagenesis and RNAi with genomic eDNA hybrids. I will characterize all resulting phenotypes and use these strains to identify modifiers of MeCP2 and/or CG10042 through an F 1 modifier screen to gain insight into pathways of MeCP2 function, which until now has been studied only in vitro. Through the proposed genetic studies, I hope to identify proteins that can be then studied to investigate the mechanisms of the mental and neurological dysfunction in Rett and related disorders.