Zhang Z, Shibahara K, Stillman B
Cold Spring Harbor Laboratory, New York 11724, USA.
Nature. 2000 Nov 9;408(6809):221-5. doi: 10.1038/35041601.
Formation of a heterochromatin-like structure results in transcriptional silencing at the HM mating-type loci and telomeres in Saccharomyces cerevisiae. Once formed, such epigenetically determined structures are inherited for many mitotic divisions. Here we show that mutations in the proliferating cell nuclear antigen (PCNA), an essential component at the DNA replication fork, reduced repression of genes near a telomere and at the silent mating-typelocus, HMR. The pol30-8 mutant displayed coexistence of both repressed (pink) and de-repressed (white) cells within a single colony when assayed with the ADE2 gene inserted at HMR. Unlike pol30-8, the pol30-6 and pol30-79 mutants partially reduced gene silencing at telomeres and the HMR and synergistically decreased silencing in cells lacking chromatin assembly factor 1 (CAF-1). All silencing defective mutants showed reduced binding to CAF-1 in vitro and altered chromatin association of the CAF-1 large subunit in vivo. Thus, PCNA participates in inheritance of both DNA and epigenetic chromatin structures during the S phase of the cell cycle, the latter by at least two mechanisms.
异染色质样结构的形成导致酿酒酵母中HM交配型基因座和端粒处的转录沉默。一旦形成,这种由表观遗传决定的结构会在许多有丝分裂过程中遗传。我们在此表明,增殖细胞核抗原(PCNA)是DNA复制叉的一个重要组成部分,其突变会降低端粒附近基因以及沉默交配型基因座HMR处基因的抑制作用。当用插入HMR的ADE2基因进行检测时,pol30 - 8突变体在单个菌落中表现出受抑制(粉红色)和去抑制(白色)细胞的共存。与pol30 - 8不同,pol30 - 6和pol30 - 79突变体部分降低了端粒和HMR处的基因沉默,并在缺乏染色质组装因子1(CAF - 1)的细胞中协同降低了沉默作用。所有沉默缺陷突变体在体外与CAF - 1的结合减少,并且在体内CAF - 1大亚基的染色质结合发生改变。因此,PCNA在细胞周期的S期参与DNA和表观遗传染色质结构的遗传,后者至少通过两种机制实现。