Interaction of Bacteroides fragilis pLV22a relaxase and transfer DNA with Escherichia coli RP4-TraG coupling protein.
Interaction of Bacteroides fragilis pLV22a relaxase and transfer DNA with Escherichia coli RP4-TraG coupling protein.
复制标题
脆弱拟杆菌 pLV22a 松弛酶和转移 DNA 与大肠杆菌 RP4-TraG 偶联蛋白的相互作用。
DOI：
10.1111/j.1365-2958.2007.05967.x
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发表时间：
2007
影响因子：
3.6
通讯作者：
Hecht,DavidW
中科院分区：
文献类型：
--
作者：
Thomas,Johnson;Hecht,DavidW
当与 IncP 接合质粒 RP4 共存时，许多拟杆菌转移因子在大肠杆菌中是可移动的，但不是 F。为了开始表征拟杆菌可移动转移因子和 RP4 交配通道之间的潜在相互作用，我们对 RP4 的 DNA 加工 (dtr)、交配对形成 (mpf) 和 traG 偶联基因进行了突变和删除。 测试拟杆菌质粒pLV22a的动员。 pLV22a 的动员需要所有 10mpf，但没有四个基因。 RP4 TraG 偶联蛋白 (CP) 也是 pLV22a 动员所必需的，但可以被 F TraD CP 的 C 端缺失突变体取代。评估了 TraG CP 与松弛酶蛋白以及 RP4 和 pLV22a 的转移 DNA 的潜在相互作用。叠加分析鉴定了 TraG 与分别来自 pLV22a 和 RP4 的 MbpB 和 TraI 的松弛酶蛋白之间的有效相互作用。农杆菌转移免疫沉淀 (TrIP) 测定还鉴定了 TraG 与 RP4 和 pLV22a 转移 DNA 之间的相互作用。因此，拟杆菌LV22a在大肠杆菌中的动员。 大肠杆菌利用 RP4 Mpf 和 CP 功能，包括松弛体和 RP4 CP 之间的相互作用，类似于同源 RP4 质粒。
ManyBacteroidestransfer factors are mobilizable inEscherichia coliwhen coresident with the IncP conjugative plasmid RP4, but not F. To begin characterization and potential interaction betweenBacteroidesmobilizable transfer factors and the RP4 mating channel, both mutants and deletions of the DNA processing (dtr), mating pair formation (mpf) andtraGcoupling genes of RP4 were tested for mobilization ofBacteroidesplasmid pLV22a. All 10mpfbut none of the fourdtrgenes were required for mobilization of pLV22a. The RP4 TraG coupling protein (CP) was also required for mobilization of pLV22a, but could be substituted by a C‐terminal deletion mutant of the F TraD CP. Potential interactions of the TraG CP with relaxase protein(s) and transfer DNA of both RP4 and pLV22a were assessed. Overlay assays identified productive interactions between TraG and the relaxase proteins of both MbpB and TraI from pLV22a and RP4 respectively. TheAgrobacteriumTransfer‐ImmunoPrecipitation (TrIP) assay also identified an interaction between TraG and both RP4 and pLV22a transfer DNA. Thus, mobilization of theBacteroidespLV22a inE. coliutilizes both RP4 Mpf and CP functions including an interaction between the relaxosome and the RP4 CP similar to that of cognate RP4 plasmid.