REGULATION OF THE NEURAL DETERMINATION GENE MASH1

描述：该应用程序的重点是调节和功能 小鼠前神经基因MASH-1。MASH-1编码DNA结合蛋白 的bHLH家族;其表达和功能表型的丧失， 被广泛研究。 在上一个授予期间， 申请人已着手确定顺式和交易因素 控制小鼠神经系统中MASH-1的表达。 这些 经过努力，鉴定出了一个1.2kB的序列， 驱动lacZ报告基因的模式类似于 内源性MASH-1表达。 相关顺式作用的位置 删除结构进一步缩小了元素;三个 在1.2kB片段内鉴定出150 bpp区域，其缺失导致 部分lacZ表达的显著缺失。 也是 发现报告基因构建体的表达在 纯合MASH-1突变背景，表明MASH-1蛋白可能 作为其自身表达的抑制剂。 最后，申请人有 提供了令人信服的初步证据，与先前相反， 据报道，MASH-1基因敲除小鼠在CNS中显示出显著的缺陷。 旨在 1-3该提案的目的是确定特定的结合位点， 1.2kB MASH-1突变体大脑中的转录因子。

DESCRIPTION: This application focusses on regulation and function of the mouse proneural gene MASH-1. MASH-1 encodes a DNA binding protein of the bHLH family; its expression and loss of function phenotype has been studied extensively. During the previous granting period the applicant has set out to identify cis- and transacting factors controlling MASH-1 expression in the mouse nervous system. These efforts resulted in the identification of a 1.2kB sequence which is able to drive a lacZ reporter gene in a pattern similar to that one of endogenous MASH-1 expression. The location of relevant cis-acting elements has been further narrowed down by deletion constructs; three 150bpp regions were identified within the 1.2kB piece whose deletion led to significant losses of part of the lacZ expression. It was also discovered that expression of the reporter constructs is upregulated in homozygous MASH-1 mutant background, suggesting that MASH-1 protein may act as an inhibitor of its own expression. Finally, the applicant has furnished convincing preliminary evidence that, contrary to prior reports, MASH-1 knockout mice show significant defects in the CNS. Aims 1-3 of this proposal intend to identify specific binding sites for transcription factors in the 1.2kB MASH-1 mutant brains.