CTnDOT integrase interactions with attachment site DNA and control of directionality of the recombination reaction.
CTnDOT integrase interactions with attachment site DNA and control of directionality of the recombination reaction.
复制标题
CTnDOT 整合酶与附着位点 DNA 相互作用并控制重组反应的方向性。
DOI：
10.1128/jb.00351-10
复制
发表时间：
2010
影响因子：
3.2
通讯作者：
Gardner,JeffreyF
中科院分区：
文献类型：
--
作者：
Wood,MargaretM;Dichiara,JeanneM;Yoneji,Sumiko;Gardner,JeffreyF
IntDOT is a tyrosine recombinase encoded by the conjugative transposon CTnDOT. The core binding (CB) and catalytic (CAT) domains of IntDOT interact with core-type sites adjacent to the regions of strand exchange, while the N-terminal arm binding (N) domain interacts with arm-type sites distal to the core. Previous footprinting experiments identified five arm-type sites, but how the arm-type sites participate in the integration and excision of CTnDOT was not known.In vitrointegration assays with substrates containing arm-type site mutants demonstrated thatattDOTsequences containing mutations in the L1 arm-type site or in the R1 and R2 or R1 and R2′ arm-type sites were dramatically defective in integration. Substrates containing mutations in the L1 and R1 arm-type sites showed a 10- to 20-fold decrease in detectablein vitroexcision, but introduction of multiple arm-type site mutations inattRdid not have an effect on the excision frequency. A sixth arm-type site, the R1′ site, was also identified and shown to be required for integration and important for efficient excision. These results suggest that intramolecular IntDOT interactions are required for integration, while the actions of accessory factors are more important for excision. Gel shift assays performed in the presence of core- and arm-type site DNAs showed that IntDOT affinity for theattDOTcore was enhanced when IntDOT was simultaneously bound to arm-type site DNA.
