Jayaram M, Sutton A, Broach J R
Mol Cell Biol. 1985 Sep;5(9):2466-75. doi: 10.1128/mcb.5.9.2466-2475.1985.
Stable propagation of the yeast plasmid 2 microns requires an origin of replication, a cis-active locus designated REP3, and two plasmid-encoded proteins which are the products of the REP1 and REP2 genes. The three REP loci appear to constitute a partitioning system, ensuring equal distribution of plasmid molecules to mother and daughter cells after mitosis. We have localized the REP3 site completely within a segment of five-and-one-half direct tandem repeats of a 62-base-pair unit, bordered by HpaI and AvaI restriction sites within the large unique region of the 2 microns genome. In addition, we find that the repeated elements are functionally distinct. Only a subset of the repeats is necessary to promote full partitioning activity. The other repeats appear to promote plasmid transcription. These results are discussed in the context of a model of plasmid copy control involving titration of a plasmid-specific protein by the repeated elements within REP3.
酵母质粒2μm的稳定复制需要一个复制起点、一个名为REP3的顺式作用位点以及两种由质粒编码的蛋白质,即REP1和REP2基因的产物。这三个REP位点似乎构成了一个分配系统,确保有丝分裂后质粒分子能平均分配到母细胞和子细胞中。我们已将REP3位点完全定位在一个62个碱基对单位的五点五个直接串联重复序列片段内,该片段由2μm基因组大的单一区域内的HpaI和AvaI限制酶切位点界定。此外,我们发现这些重复元件在功能上是不同的。只有一部分重复序列对于促进完全分配活性是必需的。其他重复序列似乎促进质粒转录。这些结果将在一个质粒复制控制模型的背景下进行讨论,该模型涉及通过REP3内的重复元件对一种质粒特异性蛋白质进行滴定。