Mutations in the 2-microns circle site-specific recombinase that abolish recombination without affecting substrate recognition.
Mutations in the 2-microns circle site-specific recombinase that abolish recombination without affecting substrate recognition.
复制标题
2 微米圆位点特异性重组酶发生突变，可消除重组而不影响底物识别。
DOI：
10.1073/pnas.84.8.2189
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发表时间：
1987
影响因子：
11.1
通讯作者：
Makkuni Jayaram
中科院分区：
文献类型：
--
作者：
P. Prasad;L. Young;Makkuni Jayaram
由酵母质粒2微米环（FLP）编码的位点特异性重组酶通过酪氨酸残基与其底物DNA形成瞬时共价连接，所述酪氨酸残基似乎位于其COOH末端附近。FLP的COOH末端与重组酶Int家族的COOH末端的同源性表明FLP的酪氨酸-343可能参与形成DNA-蛋白质桥。我们把酪氨酸-343突变成苯丙氨酸或丝氨酸。我们证明，结合的两个突变体蛋白质的每一个其基板是无法区分的野生型FLP。然而，这两种突变蛋白都不能催化链切割和重组。
The site-specific recombinase encoded by the yeast plasmid 2-microns circle (FLP) forms a transient covalent linkage with its substrate DNA via a tyrosine residue, which appears to be located near its COOH terminus. The homology of the COOH terminus of FLP with that of the Int family of recombinases suggests that tyrosine-343 of FLP could be involved in forming the DNA-protein bridge. We have mutated tyrosine-343 to a phenylalanine or serine. We demonstrate that the binding of each of the two mutant proteins to its substrate is indistinguishable from that of wild-type FLP. However, both mutant proteins are incapable of catalyzing strand cleavage and recombination.