Greene Eric C, Mizuuchi Kiyoshi
Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
Mol Cell. 2002 Dec;10(6):1367-78. doi: 10.1016/s1097-2765(02)00733-5.
The Mu transpososome can distinguish between proximal and distal DNA during the selection of a site for transposition. This phenomenon, termed target immunity, involves MuA-stimulated removal of MuB oligomers from sites near the Mu genome. Using a combination of ensemble and single-molecule fluorescence methods, we show that the MuA tetramer can stably associate with the DNA-bound MuB oligomer and is more efficient than monomeric MuA at stimulating the dissociation of MuB from DNA. In addition, we demonstrate that DNA looping is essential for efficient disassembly of the MuB oligomer. We propose a model in which the MuA tetramer forms a multivalent complex with the MuB oligomer and catalyzes the processive removal of MuB from DNA.
在选择转座位点的过程中,Mu转座体能够区分近端和远端DNA。这种现象被称为靶标免疫,涉及MuA刺激下从Mu基因组附近位点去除MuB寡聚体。通过结合整体和单分子荧光方法,我们表明MuA四聚体可以与结合DNA的MuB寡聚体稳定结合,并且在刺激MuB从DNA上解离方面比单体MuA更有效。此外,我们证明DNA环化对于MuB寡聚体的有效拆解至关重要。我们提出了一个模型,其中MuA四聚体与MuB寡聚体形成多价复合物,并催化MuB从DNA上的逐步去除。