Covitz P A, Mitchell A P
Department of Microbiology, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
Genes Dev. 1993 Aug;7(8):1598-608. doi: 10.1101/gad.7.8.1598.
The RME1 gene product, a negative regulator of meiosis with three zinc finger motifs, acts by preventing transcript accumulation from IME1, whose product is required for meiotic gene expression. We have isolated a 404-bp segment from a region 2 kb upstream of IME1 that is sufficient for RME1-dependent repression of a heterologous promoter. This DNA contains an RME1-response element (RRE) and another region called the modulation region. The modulation region is required for repression because DNA containing the RRE alone did not repress but was able to confer RME1-dependent transcriptional activation of a reporter gene. In gel mobility retardation assays, RME1 formed a specific complex with the RRE, and RRE point mutations that reduced the affinity for RME1 also blocked repression and activation. Footprinting of the RME1-RRE complex revealed a 21-bp protected region that included the positions of these RRE mutations. We conclude that RME1 binding to this RRE is required for repression. Thus, the mechanism of meiotic inhibition by RME1 is direct transcriptional repression of IME1.
RME1基因产物是一种具有三个锌指基序的减数分裂负调控因子,其作用机制是阻止IME1的转录积累,而IME1的产物是减数分裂基因表达所必需的。我们从IME1上游2 kb区域分离出一个404 bp的片段,该片段足以实现RME1对异源启动子的抑制作用。这段DNA包含一个RME1反应元件(RRE)和另一个称为调节区域的区域。调节区域对于抑制作用是必需的,因为仅包含RRE的DNA不会产生抑制作用,但能够赋予报告基因RME1依赖的转录激活。在凝胶迁移率阻滞分析中,RME1与RRE形成了一个特异性复合物,降低对RME1亲和力的RRE点突变也会阻断抑制作用和激活作用。对RME1-RRE复合物的足迹分析揭示了一个21 bp的保护区域,该区域包含这些RRE突变的位置。我们得出结论,RME1与该RRE的结合是抑制作用所必需的。因此,RME1对减数分裂的抑制机制是对IME1的直接转录抑制。