Hayes F, Davis M A, Austin S J
Laboratory of Chromosome Biology, ABL-Basic Research Program, NCI-Frederick Cancer Research and Development Center, Maryland 21702.
J Bacteriol. 1993 Jun;175(11):3443-51. doi: 10.1128/jb.175.11.3443-3451.1993.
The par region of bacteriophage P7 is responsible for active partition of the P7 plasmid prophage into daughter cells. The cis-acting partition site was defined precisely as a 75-bp sequence that was necessary and sufficient to promote correct segregation of an unstable vector plasmid when the two P7 partition proteins, ParA and ParB, were supplied in trans. Roughly the same region was necessary to exert partition-mediated incompatibility. The minimal site contains an integration host factor (IHF) protein binding site bracketed by regions containing heptamer repeat sequences that individually bind ParB. An additional sequence forms the left boundary of the site. Site-directed mutations in the latter sequence, as well as the IHF motif and the rightmost ParB box, blocked site function. Although the P7 site shares 55% sequence identity with its counterpart in bacteriophage P1, functional interactions between the partition sites and the Par proteins of the two plasmids were entirely species specific in vivo. The P1 sequence has similar IHF and ParB binding motifs, but the left boundary sequence differs radically and may define a point of species-specific contact with the Par proteins. No evidence was found for the existence of a functional P7 analog of the P1 parS core, a small subregion of the P1 site that, in isolation, acts as an enfeebled partition site with modified incompatibility properties.
噬菌体P7的par区域负责将P7质粒原噬菌体主动分配到子代细胞中。顺式作用分配位点被精确地定义为一个75碱基对的序列,当两个P7分配蛋白ParA和ParB以反式提供时,该序列对于促进不稳定载体质粒的正确分离是必要且充分的。大致相同的区域对于发挥分配介导的不相容性也是必需的。最小位点包含一个整合宿主因子(IHF)蛋白结合位点,其两侧是含有七聚体重复序列的区域,这些区域分别结合ParB。一个额外的序列构成了该位点的左边界。该序列以及IHF基序和最右侧的ParB框中的定点突变均阻断了位点功能。尽管P7位点与其在噬菌体P1中的对应位点具有55%的序列同一性,但在体内,两个质粒的分配位点与Par蛋白之间的功能相互作用完全是物种特异性的。P1序列具有相似的IHF和ParB结合基序,但左边界序列有很大差异,可能定义了与Par蛋白物种特异性接触的点。未发现存在P1 parS核心的功能性P7类似物的证据,P1 parS核心是P1位点的一个小亚区域,单独存在时作为一个具有改变的不相容特性的弱化分配位点。