TraY and integration host factor oriT binding sites and F conjugal transfer: sequence variations, but not altered spacing, are tolerated.
TraY and integration host factor oriT binding sites and F conjugal transfer: sequence variations, but not altered spacing, are tolerated.
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TraY 和整合宿主因子 oriT 结合位点和 F 接合转移：可以容忍序列变异，但不能改变间距。
DOI：
10.1128/jb.01783-06
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发表时间：
2007
影响因子：
3.2
通讯作者：
Schildbach,JoelF
中科院分区：
文献类型：
--
作者：
Williams,SarahL;Schildbach,JoelF
细菌接合是接合质粒的单链从供体转移到受体的过程。对于F质粒，TraI是一种松弛酶或切口酶，在其特异性转移起点（oriT）结合位点sbi结合单个质粒DNA链，并在称为nic的位点切割。体外研究表明，TraI被其辅助蛋白TraY和整合宿主因子（IHF）募集到体内。TraY和IHF分别结合AandhfA的保守位点，并使DNA弯曲。由此产生的构象变化可能会传播紧张性，产生TraI可以结合的单链区域。先前由其他人进行的缺失研究表明，随着越来越长的片段（最终包含AandihfA）被缺失，含有ForiT的质粒的转移效率逐渐降低。在这里，我们描述了我们的努力，以更精确地定义的作用，通过检查多个碱基取代atsbyAandihfA的结合和质粒动员的影响。虽然我们观察到体外DNA结合亲和力的显着下降，我们看到质粒动员的影响很小，即使当AandihfA变体组合。相反，当半或全螺旋圈之间插入松弛体蛋白结合位点，动员显着减少，在某些情况下，低于检测限的测定。这些结果与AandihfA识别TraY和IHF的序列特异性有限以及松弛体蛋白相互之间需要适当的间隔和方向一致。
Bacterial conjugation is the process by which a single strand of a conjugative plasmid is transferred from donor to recipient. For F plasmid, TraI, a relaxase or nickase, binds a single plasmid DNA strand at its specific origin of transfer (oriT) binding site,sbi, and cleaves at a site callednic. In vitro studies suggest TraI is recruited tosbiby its accessory proteins, TraY and integration host factor (IHF). TraY and IHF bind conservedoriTsitessbyAandihfA, respectively, and bend DNA. The resulting conformational changes may propagate tonic, generating the single-stranded region that TraI can bind. Previous deletion studies performed by others showed transfer efficiency of a plasmid containing ForiTdecreased progressively as increasingly longer segments, ultimately containing bothsbyAandihfA, were deleted. Here we describe our efforts to more precisely define the role ofsbyAandihfAby examining the effects of multiple base substitutions atsbyAandihfAon binding and plasmid mobilization. While we observed significant decreases in in vitro DNA-binding affinities, we saw little effect on plasmid mobilization even whensbyAandihfAvariants were combined. In contrast, when half or full helical turns were inserted between the relaxosome protein-binding sites, mobilization was dramatically reduced, in some cases below the detectable limit of the assay. These results are consistent with TraY and IHF recognizingsbyAandihfAwith limited sequence specificity and with relaxosome proteins requiring proper spacing and orientation with respect to each other.
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DOI：
--
发表时间：
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期刊：
Transactions of the Association of American Physicians
影响因子：
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影响因子：
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影响因子：
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作者：
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通讯作者：
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