NUCLEASE HYPERSENSITIVE REGIONS WITH ADJACENT POSITIONED NUCLEOSOMES MARK THE GENE BOUNDARIES OF THE PHO5/PHO3 LOCUS IN YEAST
NUCLEASE HYPERSENSITIVE REGIONS WITH ADJACENT POSITIONED NUCLEOSOMES MARK THE GENE BOUNDARIES OF THE PHO5/PHO3 LOCUS IN YEAST
复制标题
DOI：
10.1002/j.1460-2075.1986.tb04551.x
复制
发表时间：
1986-10-01
期刊：
影响因子：
11.4
通讯作者：
HORZ, W
中科院分区：
文献类型：
--
作者：
ALMER, A;HORZ, W
分析了两个串联酸性磷酸酶基因（PHO 5和PHO 3）从酿酒酵母的染色质结构的条件下，强烈调控PHO 5基因被抑制。用DNA酶I、DNA酶II、微球菌核酸酶和限制性核酸酶的消化实验揭示了PHO 5/PHO 3位点处存在5个超敏位点，其中两个在PHO 5上游，距离为920和370 bp，一个在两个基因之间，两个在PHO 3下游。 如间接末端标记实验所示，特定定位的核小体位于这些超敏感位点旁边。从这些实验中推导出的位置可以通过监测各种限制性位点对相应核酸酶的可接近性来验证。推定的连接区域内的网站约50-60%敏感，而位于核小体核心内的网站是耐药的。将微球菌核酸酶抑制剂与两个上游超敏位点之间的探针杂交，导致原本规则的核小体DNA模式中断。这直接表明，这些超敏感位点代表有序核小体阵列中的间隙。
The chromatin structure of two tandemly linked acid phosphatase genes (PHO5 and PHO3) from Saccharomyces cerevisiae was analyzed under conditions at which the strongly regulated PHO5 gene is repressed. Digestion experiments with DNase I, DNase II, micrococcal nuclease and restriction nucleases reveal the presence of five hypersensitive sites at the PHO5/PHO3 locus, two of them upstream of PHO5 at distances of 920 and 370 bp, one in between the two genes and two downstream of PHO3. Specifically positioned nucleosomes are located next to these hypersensitive sites as shown by indirect end-labeling experiments. The positions deduced from these experiments could be verified by monitoring the accessibility of various restriction sites to the respective nucleases. Sites within putative linker regions were about 50-60% susceptible, whereas sites located within nucleosome cores were resistant. Hybridizing micrococcal nuclease digests to a probe from in between the two upstream hypersensitive sites leads to an interruption of an otherwise regular nucleosomal DNA pattern. This shows directly that these hypersensitive sites represents gaps within ordered nucleosomal arrays.