Martin K A, Friedman S A, Austin S J
Laboratory of Chromosome Biology, National Cancer Institute-Frederick Cancer Research Facility, MD 21701.
Proc Natl Acad Sci U S A. 1987 Dec;84(23):8544-7. doi: 10.1073/pnas.84.23.8544.
We have defined a minimal partition site, parS, from the plasmid P1. It contains sufficient cis-acting information to direct accurate segregation of low-copy-number plasmids that contain it as long as the two essential P1 Par proteins are supplied in trans. The site is, at most, 34 base pairs and contains a perfect 13-base-pair inverted repeat. Site-directed mutations were made within the repeat sequence that abolished activity whether or not the symmetry of the palindrome was maintained. Partition appears to be a competitive process, as differentially marked plasmids carrying the same type of partition site are not independently segregated but are randomly distributed with respect to each other. We have studied competition between plasmids carrying various fragments encompassing the parS site. As expected, two plasmids carrying the minimal parS site compete with each other. However, a sequence that lies to the left of the minimal parS site acts as a major modulator of this competition, changing the specificity of the competitive effect completely. Thus, this adjacent sequence appears to be an important determinant of the specificity of the wild-type P1 partition system without being necessary for its efficient function.
我们从质粒P1中定义了一个最小分配位点parS。它包含足够的顺式作用信息,只要以反式提供两种必需的P1 Par蛋白,就能指导含有它的低拷贝数质粒进行精确分离。该位点最多34个碱基对,包含一个完美的13个碱基对的反向重复序列。在重复序列内进行了定点突变,无论回文结构的对称性是否保持,这些突变都会消除活性。分配似乎是一个竞争性过程,因为携带相同类型分配位点的不同标记质粒不是独立分离的,而是彼此随机分布。我们研究了携带包含parS位点的各种片段的质粒之间的竞争。正如预期的那样,两个携带最小parS位点的质粒相互竞争。然而,位于最小parS位点左侧的一个序列是这种竞争的主要调节因子,它完全改变了竞争效应的特异性。因此,这个相邻序列似乎是野生型P1分配系统特异性的一个重要决定因素,但其高效功能并非必需。