Synapsis and catalysis by activated Tn3 resolvase mutants.
Synapsis and catalysis by activated Tn3 resolvase mutants.
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DOI：
10.1093/nar/gkn885
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发表时间：
2008-12
影响因子：
14.9
通讯作者：
Stark WM
中科院分区：
文献类型：
--
作者：
Olorunniji FJ;He J;Wenwieser SV;Boocock MR;Stark WM
丝氨酸重组酶Tn3分解酶催化两个114 bp位点之间的重组，每个位点包含三个分解酶二聚体的结合位点。我们分析了具有“激活”突变的解溶变异体的体外特性，当res的其余部分缺失时，它可以催化res的结合位点I的重组。由这些变异促进的Site I × Site I重组与野生型分解促进的res × res重组一样快。激活的变体具有较低的拓扑选择性，并且不再需要亚基之间的2-3 '界面，而这对于野生型分解酶介导的重组是必不可少的。它们还促进稳定突触的形成，包括一个分解四聚体和两个位点i的拷贝。被激活的突变体对DNA链的切割相对于突触的速率是缓慢的。在没有DNA的情况下，没有检测到稳定的分解四聚体或与单个位点I结合。我们的结果使我们得出结论，突触是通过分解单体与位点I的顺序结合，然后是两个位点I二聚体复合物的相互作用来组装的。我们讨论了我们的结果对突触机制和野生型分解重组调控的影响。
The serine recombinase Tn3 resolvase catalyses recombination between two 114 bp res sites, each of which contains binding sites for three resolvase dimers. We have analysed the in vitro properties of resolvase variants with ‘activating’ mutations, which can catalyse recombination at binding site I of res when the rest of res is absent. Site I × site I recombination promoted by these variants can be as fast as res × res recombination promoted by wild-type resolvase. Activated variants have reduced topological selectivity and no longer require the 2–3′ interface between subunits that is essential for wild-type resolvase-mediated recombination. They also promote formation of a stable synapse comprising a resolvase tetramer and two copies of site I. Cleavage of the DNA strands by the activated mutants is slow relative to the rate of synapsis. Stable resolvase tetramers were not detected in the absence of DNA or bound to a single site I. Our results lead us to conclude that the synapse is assembled by sequential binding of resolvase monomers to site I followed by interaction of two site I-dimer complexes. We discuss the implications of our results for the mechanisms of synapsis and regulation in recombination by wild-type resolvase.
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影响因子：
16
作者：
Mouw, Kent W.;Rowland, Sally-J.;Gajjar, Mark M.;Boocock, Martin R.;Stark, W. Marshall;Rice, Phoebe A.
通讯作者：
Rice, Phoebe A.
影响因子：
11.4
作者：
KLIPPEL, A;KANAAR, R;COZZARELLI, NR
通讯作者：
COZZARELLI, NR
DOI：
10.1073/pnas.1533177100
发表时间：
2003-07-22
影响因子：
11.1
作者：
Akopian, A;He, JY;Stark, WM
通讯作者：
Stark, WM
影响因子：
64.5
作者：
REED, RR;GRINDLEY, NDF
通讯作者：
GRINDLEY, NDF
影响因子：
11.4
作者：
RICE, PA;STEITZ, TA
通讯作者：
STEITZ, TA