Deletion mutations affecting autonomously replicating sequence ARS1 of Saccharomyces cerevisiae.
Deletion mutations affecting autonomously replicating sequence ARS1 of Saccharomyces cerevisiae.
复制标题
影响酿酒酵母自主复制序列 ARS1 的缺失突变。
DOI：
10.1128/mcb.4.11.2455-2466.1984
复制
发表时间：
1984
影响因子：
5.3
通讯作者：
Campbell,JL
中科院分区：
文献类型：
--
作者：
Celniker,SE;Sweder,K;Srienc,F;Bailey,JE;Campbell,JL
含有特定的酵母染色体序列的DNA被称为ARSstrans，这些DNA转化酿酒酵母的频率很高，当被导入S时，可以作为质粒在染色体外复制。通过变换酿酒。以确定自主复制INS所需的最小序列的边界。Cerevisiae，我们已经对第一个染色体ARS进行了体外诱变。我们在ARS1中发现了三个不同的功能结构域，而不是确定一个明显且连续的片段来调节ARS+表型。我们将结构域A定义为11个碱基对(BP)序列，该序列也在大多数其他AR区域中发现。对于高频变换来说，这是必要的，但不是充分的。结构域B不能介导高频转化或自身复制，是高效、稳定复制含有结构域A的质粒所必需的，正如我们所定义的那样，结构域B在ARS1中与结构域A连续，但在两者之间插入4bp不影响复制。B结构域的大小上限为109个碱基，下限为46个碱基。ARS1的B结构域与其他任何ARS序列之间没有明显的序列同源性。最后，根据我们的删除，C结构域被定义为A结构域与B结构域相反侧至少200bp的侧翼，也是A结构域INS稳定性所必需的。酿酒。只有在没有B结构域的情况下，才能观察到结构域C的缺失的影响，至少通过本研究中使用的分析方法可以观察到，并讨论了这一发现的意义。
DNAs that contain specific yeast chromosomal sequences calledARSstransformSaccharomyces cerevisiaeat high frequency and can replicate extrachromosomally as plasmids when introduced intoS. cerevisiaeby transformation. To determine the boundaries of the minimal sequences required for autonomous replication inS. cerevisiae, we have carried out in vitro mutagenesis of the first chromosomalARSdescribed,ARS1. Rather than identifying a distinct and continuous segment that mediates theARS+phenotype, we find three different functional domains withinARS1. We define domain A as the 11-base-pair (bp) sequence that is also found at most otherARSregions. It is necessary but not sufficient for high-frequency transformation. Domain B, which cannot mediate high-frequency transformation, or replicate by itself, is required for efficient, stable replication of plasmids containing domain A. Domain B, as we define it, is continuous with domain A inARS1, but insertions of 4 bp between the two do not affect replication. The extent of domain B has an upper limit of 109 bp and a lower limit of 46 bp in size. There is no obvious sequence homology between domain B ofARS1and any otherARSsequence. Finally, domain C is defined on the basis of our deletions as at least 200 bp flanking domain A on the opposite side from domain B and is also required for the stability of domain A inS. cerevisiae. The effect of deletions of domain C can be observed only in the absence of domain B, at least by the assays used in the current study, and the significance of this finding is discussed.