Kaykov Atanas, Arcangioli Benoit
Unité de dynamique du Génome, Unités de Recherche Associées, 1644 du Centre National de la Recherche Scientifique, Département de la Structure et Dynamique des Génomes, Institut Pasteur, 75724 Paris 15, France.
Curr Biol. 2004 Nov 9;14(21):1924-8. doi: 10.1016/j.cub.2004.10.026.
The sexual locus mat1, in the fission yeast Schizosaccharomyces pombe, efficiently switches between the two mating types, P and M, by a process similar to gene conversion, using the silent mat2-P and mat3-M loci, respectively, as donors of the P and M genetic information . It has been proposed that an asymmetrically inherited, site- and strand-specific imprint at mat1 initiates the mating-type switching process . The molecular nature of the imprint is controversial; it was initially described as a double-strand break and then as a single-strand lesion or a strand-specific, alkali-labile modification . Here, we use E. coli DNA ligase in vitro to demonstrate that the imprint is a nick with no resection of nucleotides. By using ligation-mediated PCR, we show that the nick contains 3'OH and 5'OH unphosphorylated termini resistant to RNase treatments. This nonmutational mark on one of the DNA strands provides the first example of a novel type of imprint.
在裂殖酵母粟酒裂殖酵母中,性位点mat1通过类似于基因转换的过程,分别利用沉默的mat2 - P和mat3 - M位点作为P和M遗传信息的供体,在两种交配型P和M之间高效切换。有人提出,mat1处不对称遗传的、位点和链特异性印记启动了交配型转换过程。印记的分子本质存在争议;它最初被描述为双链断裂,然后被描述为单链损伤或链特异性的、碱不稳定修饰。在这里,我们在体外使用大肠杆菌DNA连接酶来证明印记是一个没有核苷酸切除的切口。通过使用连接介导的PCR,我们表明该切口含有对RNase处理有抗性的3'OH和5'OH未磷酸化末端。DNA一条链上的这种非突变标记提供了一种新型印记的首个例子。