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首页> 外文期刊>Molecular and Cellular Biology >Histone H1 expressed in Saccharomyces cerevisiae binds to chromatin and affects survival, growth, transcription, and plasmid stability but does not change nucleosomal spacing.
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Histone H1 expressed in Saccharomyces cerevisiae binds to chromatin and affects survival, growth, transcription, and plasmid stability but does not change nucleosomal spacing.

机译：在酿酒酵母中表达的组蛋白H1与染色质结合并影响存活，生长，转录和质粒稳定性，但不会改变核小体间距。

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Histone H1 is proposed to serve a structural role in nucleosomes and chromatin fibers, to affect the spacing of nucleosomes, and to act as a general repressor of transcription. To test these hypotheses, a gene coding for a sea urchin histone H1 was expressed from the inducible GAL1 promoter in Saccharomyces cerevisiae by use of a YEp vector for high expression levels (strain YCL7) and a centromere vector for low expression levels (strain YCL1). The H1 protein was identified by its inducibility in galactose, its apparent molecular weight, and its solubility in 5% perchloric acid. When YCL7 was shifted from glucose to galactose for more than 40 h to achieve maximal levels of H1, H1 could be copurified in approximately stoichiometric amounts with core histones of Nonidet P-40-washed nuclei and with soluble chromatin fractionated on sucrose gradients. While S. cerevisiae tolerated the expression of low levels of H1 in YCL1 without an obvious phenotype, the expression of high levels of H1 correlated with greatly reduced survival, inhibition of growth, and increased plasmid loss but no obvious change in the nucleosomal repeat length. After an initial induction, RNA levels for GAL1 and H1 were drastically reduced, suggesting that H1 acts by the repression of galactose-induced genes. Similar effects, but to a lower extent, were observed when the C-terminal tail of H1 was expressed.
机译：提出组蛋白H1在核小体和染色质纤维中起结构作用，影响核小体的间隔，并充当转录的一般阻遏物。为了检验这些假设，使用高表达水平的YEp载体（YCL7菌株）和低表达水平的着丝粒载体（YCL1菌株）在酿酒酵母中由诱导型GAL1启动子表达了编码海胆组蛋白H1的基因。 。通过在半乳糖中的可诱导性，其表观分子量以及在5％高氯酸中的溶解度来鉴定H1蛋白。当YCL7从葡萄糖变为半乳糖超过40小时以达到最大水平的H1时，可以将H1与Nonidet P-40洗涤核的核心组蛋白以可溶染色质按蔗糖梯度分级分离，以大约化学计量的量共纯化。尽管酿酒酵母可以耐受YCL1中低水平的H1表达而没有明显的表型，但高水平的H1表达与存活率降低，生长抑制和质粒损失增加相关，但核小体重复长度没有明显变化。初始诱导后，GAL1和H1的RNA水平急剧下降，表明H1通过抑制半乳糖诱导的基因起作用。表达H1的C末端尾巴时，观察到类似的效果，但程度较低。

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