Ayoub N, Goldshmidt I, Lyakhovetsky R, Cohen A
Department of Molecular Biology, The Hebrew University-Hadassah Medical School, Jerusalem, Israel.
Genetics. 2000 Nov;156(3):983-94. doi: 10.1093/genetics/156.3.983.
REII is a Schizosaccharomyces pombe repression element located at the centromere-proximal end of the mat silent domain. Here we show that inversion of REII enhances silencing on its centromere-proximal side while suppressing silencing on its centromere-distal side. Transplacement of REII to a position 2.5 kb from its native locus extends the region of stringent repression to the new REII site. These results suggest that REII defines a mat silent domain boundary by acting preferentially toward its centromere-distal side. To investigate cooperation between REII and a K-region sequence that shares homology with the centromeric dg dh repeats (cen2 homology), we targeted combinations of these elements to an ectopic site and monitored expression of an adjacent reporter gene. Centromeric dh-like sequences conferred low-level silencing on the adjacent reporter gene, and REII, which did not display silencing activity on its own, enhanced cen2 homology-mediated silencing. Cooperation was also apparent at the mat locus, where deletion of REII impaired repression stability. We propose that REII and the cen2 homology play different yet complementary roles in silencing establishment and inheritance at the mat locus.
REII是位于裂殖酵母(Schizosaccharomyces pombe)沉默结构域着丝粒近端的一种抑制元件。在此我们表明,REII的倒位增强了其着丝粒近端一侧的沉默作用,同时抑制了其着丝粒远端一侧的沉默作用。将REII转位至距其天然位点2.5 kb的位置,可将严格抑制区域扩展至新的REII位点。这些结果表明,REII通过优先作用于其着丝粒远端一侧来界定一个沉默结构域边界。为了研究REII与一个与着丝粒dg dh重复序列(cen2同源性)具有同源性的K区域序列之间的协同作用,我们将这些元件的组合靶向到一个异位位点,并监测相邻报告基因的表达。着丝粒样dh序列赋予相邻报告基因低水平的沉默作用,而自身不显示沉默活性的REII增强了cen2同源性介导的沉默作用。在mat位点也明显存在协同作用,在该位点REII的缺失损害了抑制稳定性。我们提出,REII和cen2同源性在mat位点沉默的建立和遗传中发挥不同但互补的作用。