Krebs J E, Kuo M H, Allis C D, Peterson C L
Program in Molecular Medicine and Department of Biochemistry and Molecular Biology, University of Massachusetts Medical Center, Worcester, Massachusetts 01605, USA.
Genes Dev. 1999 Jun 1;13(11):1412-21. doi: 10.1101/gad.13.11.1412.
Expression of the yeast HO gene in late G1 of the cell cycle requires the SWI/SNF chromatin remodeling complex, the Gcn5p histone acetyltransferase, and two different sequence-specific transcriptional activators, Swi5p and Swi4p/Swi6p. We have used chromatin immunoprecipitation assays to investigate the role of each of these trans-acting factors in establishing a cell cycle-regulated domain of histone acetylation surrounding the HO upstream regulatory region. We detect a approximately 1-kb domain of H3 and H4 acetylation that is established in mid-G1, prior to and independent of HO transcription, which then declines with kinetics similar to inactivation of HO. This cell cycle burst of histone acetylation requires Gcn5p, SWI/SNF, and the Swi5p activator, but occurs in the absence of the Swi4p activator. We also find that inactivation of the Sin3p/Rpd3p deacetylase complex leads to a high level of acetylation at the HO locus throughout the cell cycle. We propose a sequential model for activation of HO in which the Swi5p-dependent recruitment of the Gcn5p acetyltransferase requires chromatin remodeling events by the SWI/SNF complex.
酵母HO基因在细胞周期G1期晚期的表达需要SWI/SNF染色质重塑复合体、Gcn5p组蛋白乙酰转移酶以及两种不同的序列特异性转录激活因子Swi5p和Swi4p/Swi6p。我们利用染色质免疫沉淀试验来研究这些反式作用因子中的每一个在围绕HO上游调控区建立一个细胞周期调控的组蛋白乙酰化结构域中的作用。我们检测到一个约1kb的H3和H4乙酰化结构域,它在G1期中期建立,先于HO转录且不依赖于HO转录,然后其下降动力学与HO失活相似。这种组蛋白乙酰化的细胞周期爆发需要Gcn5p、SWI/SNF和Swi5p激活因子,但在没有Swi4p激活因子的情况下也会发生。我们还发现Sin3p/Rpd3p去乙酰化酶复合体的失活会导致HO基因座在整个细胞周期都处于高水平的乙酰化状态。我们提出了一个HO激活的顺序模型,其中Gcn5p乙酰转移酶的Swi5p依赖性募集需要SWI/SNF复合体进行染色质重塑事件。