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（MeCP 2）的基因突变导致Rett综合征（RTT）和男性和女性的许多其他精神发育迟滞综合征。RTT是由MECP 2的功能丧失突变引起的，MECP 2与靶DNA结合并募集Sin 3A和组蛋白脱乙酰酶以沉默转录。MeCP 2是一个更大的基因家族的一部分，甲基-CpG结合结构域（MBD）家族，这是保守的从苍蝇到人类。使用UAS-GAL 4系统在果蝇中过表达MECP 2产生表型，可能是通过与MBD基因竞争。我建议继续对全长品系进行表征，并创建额外的转基因果蝇：两个品系过度表达Rett患者中常见的MECP 2突变，一个品系产生不含MBD区域的MeCP 2。其次，我将描述果蝇MBD基因CG 10042的天然RNA和蛋白质表达模式。然后，我将通过P元件突变和RNAi基因组eDNA杂交来研究其功能。我将表征所有得到的表型，并使用这些菌株通过F1修饰剂筛选来鉴定MeCP 2和/或CG 10042的修饰剂，以深入了解MeCP 2功能的途径，到目前为止仅在体外研究。通过拟议的遗传学研究，我希望能够识别出可以研究的蛋白质，以研究Rett和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.