Lombardi Laura M, Davis Matthew D, Rine Jasper
Department of Molecular and Cell Biology, and California Institute for Quantitative Biosciences, University of California, Berkeley, California 94720.
Center for Systems and Synthetic Biology, Institute for Cellular and Molecular Biology, The University of Texas, Austin, Texas, 78712.
Genetics. 2015 Jan;199(1):105-16. doi: 10.1534/genetics.114.168039. Epub 2014 Nov 17.
The extent of chromatin compaction is a fundamental driver of nuclear metabolism . Yta7 is a chromatin-associated AAA-ATPase, the human ortholog of which, ANCCA/ATAD2 transcriptionally activates pathways of malignancy in a broad range of cancers. Yta7 directly binds histone H3, and bulk chromatin exhibits increased nucleosomal density in yta7Δ mutants. The suppression of yta7Δ mutant growth and transcriptional phenotypes in budding yeast by decreased dosage of histones H3 and H4 indicates the acute sensitivity of cells to deviations in nucleosome spacing. This study investigated the global changes in chromatin structure upon Yta7 loss or overexpression and determined which of these effects reflected direct Yta7 activity. Metagene analysis of Yta7's genome-wide localization indicated peak binding of Yta7 just downstream of the transcription start site. Cells lacking Yta7 exhibited increased nucleosome density within genes downstream of the +1 nucleosome, as defined by decreased internucleosomal distance, resulting in progressively 5'-shifted nucleosomes within the gene. In contrast, cells overexpressing Yta7 displayed profound 3'-shifts in nucleosome position and reduced nucleosome density within genes. Importantly, Yta7-bound regions were enriched for nucleosomal shifts, indicating that Yta7 acted locally to modulate nucleosome spacing. The phenotype of cells lacking both Yta7 and Rtt106, the histone H3/H4 chaperone, indicated that Yta7 functions in both Rtt106-dependent and Rtt106-independent ways to modulate nucleosome spacing within genes. This study suggested that Yta7 affected nucleosome density throughout the gene by both blocking Rtt106 from entering the gene, as shown previously at HTA1, and facilitating the loss of nucleosomes from the 5'-end.
染色质压缩程度是核代谢的一个基本驱动因素。Yta7是一种与染色质相关的AAA-ATP酶,其人类同源物ANCCA/ATAD2在多种癌症中可转录激活恶性肿瘤相关通路。Yta7直接结合组蛋白H3,并且在yta7Δ突变体中,整体染色质表现出核小体密度增加。通过降低组蛋白H3和H4的剂量来抑制芽殖酵母中yta7Δ突变体的生长和转录表型,这表明细胞对核小体间距偏差具有急性敏感性。本研究调查了Yta7缺失或过表达时染色质结构的全局变化,并确定了其中哪些效应反映了Yta7的直接活性。对Yta7全基因组定位的元基因分析表明,Yta7在转录起始位点下游有峰值结合。缺乏Yta7的细胞在+1核小体下游的基因内表现出核小体密度增加,这是由核小体间距离减小所定义的,导致基因内的核小体逐渐向5'端移动。相反,过表达Yta7的细胞在基因内的核小体位置出现了显著的3'端移动,并且核小体密度降低。重要的是,Yta7结合区域富含核小体移位,表明Yta7在局部发挥作用以调节核小体间距。同时缺乏Yta7和组蛋白H3/H4伴侣Rttl06的细胞的表型表明,Yta7以依赖Rttl06和不依赖Rttl06的方式发挥作用,以调节基因内的核小体间距。本研究表明,Yta7通过阻止Rttl06进入基因(如先前在HTA1中所示)以及促进核小体从5'端丢失,从而影响整个基因的核小体密度。