ATP hydrolysis-dependent formation of a dynamic ternary nucleoprotein complex with MutS and MutL
ATP hydrolysis-dependent formation of a dynamic ternary nucleoprotein complex with MutS and MutL
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DOI：
10.1093/nar/27.11.2325
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发表时间：
1999-06-01
影响因子：
14.9
通讯作者：
Brooks, P
中科院分区：
文献类型：
--
作者：
Galio, L;Bouquet, C;Brooks, P
大肠杆菌MutS和MutL在错配修复中的功能相互作用依赖于ATP。在这项研究中，我们表明MutS和MutL以依赖于AIP水解的方式与固定化DNA结合，并且ATP浓度接近MutS的ATP酶溶液K（m）。在去除MutS，MutL和ATP后，这个三元复合物中的大部分蛋白质不稳定结合，MutL比MutS更快地离开复合物，快速解离揭示了与蛋白质从DNA的同时快速缔合和解离的动态相互作用。表面等离子体共振分析表明，与动态结合的蛋白质相互作用的DNA比MutS-DNA复合物中的DNA更耐核酸酶消化。ATP的不可水解类似物抑制这种动态复合物的形成，但允许形成第二种类型的三元复合物，其中MutS和MutL稳定地结合到固定的DNA上。
Functional interactions of Escherichia coli MutS and MutL in mismatch repair are dependent on ATP, In this study, we show that MutS and MutL associate with immobilised DNA in a manner dependent on AIP hydrolysis and with an ATP concentration near the solution K(m) of the ATPase of MutS, After removal of MutS, MutL and ATP, much of the protein in this ternary complex is not stably associated, with MutL leaving the complex more rapidly than MutS, The rapid dissociation reveals a dynamic interaction with concurrent rapid association and dissociation of proteins from the DNA, Analysis by surface plasmon resonance showed that the DNA interacting with dynamically bound protein was more resistant to nuclease digestion than the DNA in MutS-DNA complexes. Nonhydrolysable analogs of ATP inhibit the formation of this dynamic complex, but permit formation of a second type of ternary complex with MutS and MutL stably bound to the immobilised DNA.