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酿酒酵母基因ISW2编码一种减数分裂前DNA复制所需的微管相互作用蛋白。

Saccharomyces cerevisiae gene ISW2 encodes a microtubule-interacting protein required for premeiotic DNA replication.

作者信息

Trachtulcová P, Janatová I, Kohlwein S D, Hasek J

机构信息

Institute of Microbiology, Academy of Sciences of the Czech Republic, 14220 Prague, Czech Republic.

出版信息

Yeast. 2000 Jan 15;16(1):35-47. doi: 10.1002/(SICI)1097-0061(20000115)16:1<35::AID-YEA504>3.0.CO;2-0.

Abstract

A molecular genetic characterization of the ORF YOR304W (ISW2), identified in a screen of a yeast lambdagt11 library using a monoclonal antibody that reacts with a 210 kDa mammalian microtubule-interacting protein, is presented in this paper. The protein encoded by the ORF YOR304W is 50% identical to the Drosophila nucleosome remodelling factor ISWI and is therefore a new member of the SNF2 protein family and has been recently entered into SDG as ISW2. Although not essential for vegetative growth, we found that the ISW2 gene is required for early stages in sporulation. The isw2 homozygous deletant diploid strain was blocked in the G(1) phase of the cell cycle, unable to execute the premeiotic DNA replication and progress through the nuclear meiotic division cycle. ISW2 expression from a multicopy plasmid had the same effect as deletion, but ISW2 expression from a centromeric plasmid rescued the deletion phenotype. In vegetatively growing diploid cells, the Isw2 protein was preferentially found in the cytoplasm, co-localizing with microtubules. An accumulation of the Isw2 protein within the nucleus was observed in cells entering sporulation. Together with data published very recently by Tsukiyama et al. (1999), we propose a role for the Isw2 protein in facilitating chromatin accessibility for transcriptional factor(s) that positively regulate meiosis/sporulation-specific genes.

摘要

本文介绍了对开放阅读框YOR304W(ISW2)的分子遗传学特征分析。该开放阅读框是在使用与一种210 kDa哺乳动物微管相互作用蛋白发生反应的单克隆抗体对酵母λgt11文库进行筛选时鉴定出来的。开放阅读框YOR304W编码的蛋白质与果蝇核小体重塑因子ISWI有50%的同源性,因此是SNF2蛋白家族的一个新成员,最近已作为ISW2录入酵母基因组数据库(SGD)。尽管对营养生长不是必需的,但我们发现ISW2基因在孢子形成的早期阶段是必需的。isw2纯合缺失二倍体菌株在细胞周期的G1期受阻,无法进行减数分裂前的DNA复制,也不能完成核减数分裂周期。从多拷贝质粒表达ISW2产生的效果与缺失相同,但从着丝粒质粒表达ISW2可挽救缺失表型。在营养生长的二倍体细胞中,Isw2蛋白优先存在于细胞质中,与微管共定位。在进入孢子形成的细胞中观察到Isw2蛋白在细胞核内积累。结合Tsukiyama等人(1999年)最近发表的数据,我们提出Isw2蛋白在促进染色质对正向调节减数分裂/孢子形成特异性基因的转录因子的可及性方面发挥作用。

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