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首页> 外文期刊>Journal of bacteriology >Localization Pattern of Conjugation Machinery in a Gram-Positive Bacterium
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Localization Pattern of Conjugation Machinery in a Gram-Positive Bacterium

机译：革兰氏阳性细菌中共轭机械的定位模式

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Conjugation is an efficient way for transfer of genetic information between bacteria, even between highly diverged species, and a major cause for the spreading of resistance genes. We have investigated the subcellular localization of several conserved conjugation proteins carried on plasmid pLS20 found in Bacillus subtilis. We show that VirB1, VirB4, VirB11, VirD2, and VirD4 homologs assemble at a single cell pole, but also at other sites along the cell membrane, in cells during the lag phase of growth. Bimolecular fluorescence complementation analyses showed that VirB4 and VirD4 interact at the cell pole and, less frequently, at other sites along the membrane. VirB1 and VirB11 also colocalized at the cell pole. Total internal reflection fluorescence microscopy showed that pLS20 is largely membrane associated and is frequently found at the cell pole, indicating that transfer takes place at the pole, which is a preferred site for the assembly of the active conjugation apparatus, but not the sole site. VirD2, VirB4, and VirD4 started to localize to the pole or the membrane in stationary-phase cells, and VirB1 and VirB11 were observed as foci in cells resuspended in fresh medium but no longer in cells that had entered exponential growth, although at least VirB4 was still expressed. These data reveal an unusual assembly/disassembly timing for the pLS20 conjugation machinery and suggest that specific localization of conjugation proteins in lag-phase cells and delocalization during growth are the reasons why pLS20 conjugation occurs only during early exponential phase.
机译：缀合是在细菌之间，甚至在高度分化的物种之间传递遗传信息的有效方法，并且是耐药基因传播的主要原因。我们研究了在枯草芽孢杆菌中发现的质粒pLS20上携带的几种保守的偶联蛋白的亚细胞定位。我们显示，在生长的滞后阶段，VirB1，VirB4，VirB11，VirD2和VirD4同源物在单个细胞极组装，但在沿细胞膜的其他部位也组装。双分子荧光互补分析显示，VirB4和VirD4在细胞极相互作用，而在膜的其他部位则较不频繁。 VirB1和VirB11也共定位在细胞极处。全内反射荧光显微镜检查显示，pLS20在很大程度上与膜相关，并经常在细胞极处发现，表明转移发生在极处，这是组装主动偶联装置的首选部位，但不是唯一的部位。 VirD2，VirB4和VirD4开始定位于固定相细胞中的极点或膜，并且观察到VirB1和VirB11是重悬在新鲜培养基中但不再进入指数生长的细胞中的病灶，尽管至少VirB4仍然表示。这些数据揭示了pLS20缀合机制的不寻常组装/拆卸时机，并表明在滞后期细胞中缀合蛋白的特异性定位和生长过程中的离域化是pLS20缀合仅在指数早期发生的原因。

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