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首页> 外文期刊>Molecular and Cellular Biology >Single base-pair mutations in centromere element III cause aberrant chromosome segregation in Saccharomyces cerevisiae.
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Single base-pair mutations in centromere element III cause aberrant chromosome segregation in Saccharomyces cerevisiae.

机译：着丝粒元素III中的单个碱基对突变会导致酿酒酵母中异常的染色体分离。

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摘要

In this paper we show that a 211-base pair segment of CEN3 DNA is sufficient to confer wild-type centromere function in the yeast Saccharomyces cerevisiae. We used site-directed mutagenesis of the 211-base pair fragment to examine the sequence-specific functional requirements of a conserved 11-base pair segment of centromere DNA, element III (5'-TGATTTATCCGAA-3'). Element III is the most highly conserved of the centromeric DNA sequences, differing by only a single adenine X thymine base pair among the four centromere DNAs sequenced thus far. All of the element III sequences contain specific cytosine X guanine base pairs, including a 5'-CCG-3' arrangement, which we targeted for single cytosine-to-thymine mutations by using sodium bisulfite. The effects of element III mutations on plasmid and chromosome segregation were determined by mitotic stability assays. Conversion of CCG to CTG completely abolished centromere function both in plasmids and in chromosome III, whereas conversion of CCG to TCG decreased plasmid and chromosome stability moderately. The other two guanine X cytosine base pairs in element III could be independently converted to adenine X thymine base pairs without affecting plasmid or chromosome stability. We concluded that while some specific nucleotides within the conserved element III sequence are essential for proper centromere function, other conserved nucleotides can be changed.
机译：在本文中，我们显示了CEN3 DNA的211个碱基对片段足以赋予酵母酿酒酵母野生型着丝粒功能。我们使用定点诱变的211个碱基对的片段，以检查着丝粒DNA保守的11个碱基对片段，元素III（5'-TGATTTATCCGAA-3'）的序列特异性功能需求。元素III是着丝粒DNA序列中最保守的，在迄今测序的四个着丝粒DNA中，只有一个腺嘌呤X胸腺嘧啶碱基对不同。所有元素III序列均包含特定的胞嘧啶X鸟嘌呤碱基对，包括5'-CCG-3'排列，我们使用亚硫酸氢钠靶向单胞嘧啶至胸腺嘧啶突变。通过有丝分裂稳定性测定来确定元素III突变对质粒和染色体分离的影响。 CCG向CTG的转化完全消除了质粒和III号染色体的着丝粒功能，而CCG向TCG的转化则适度降低了质粒和染色体的稳定性。元件III中的其他两个鸟嘌呤X胞嘧啶碱基对可以独立转换为腺嘌呤X胸腺嘧啶碱基对，而不会影响质粒或染色体的稳定性。我们得出的结论是，虽然保守的元素III序列中的某些特定核苷酸对于正确的着丝粒功能是必不可少的，但其他保守的核苷酸可以更改。

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