Midonet Caroline, Das Bhabatosh, Paly Evelyne, Barre Francois-Xavier
Institute for Integrative Biology of the Cell (I2BC), Université Paris Saclay, Commissariat à l'Énergie Atomique, CNRS, Université Paris Sud, 91198 Gif sur Yvette, France.
Institute for Integrative Biology of the Cell (I2BC), Université Paris Saclay, Commissariat à l'Énergie Atomique, CNRS, Université Paris Sud, 91198 Gif sur Yvette, France
Proc Natl Acad Sci U S A. 2014 Nov 25;111(47):16848-53. doi: 10.1073/pnas.1404047111. Epub 2014 Nov 10.
As in most bacteria, topological problems arising from the circularity of the two Vibrio cholerae chromosomes, chrI and chrII, are resolved by the addition of a crossover at a specific site of each chromosome, dif, by two tyrosine recombinases, XerC and XerD. The reaction is under the control of a cell division protein, FtsK, which activates the formation of a Holliday Junction (HJ) intermediate by XerD catalysis that is resolved into product by XerC catalysis. Many plasmids and phages exploit Xer recombination for dimer resolution and for integration, respectively. In all cases so far described, they rely on an alternative recombination pathway in which XerC catalyzes the formation of a HJ independently of FtsK. This is notably the case for CTXϕ, the cholera toxin phage. Here, we show that in contrast, integration of TLCϕ, a toxin-linked cryptic satellite phage that is almost always found integrated at the chrI dif site before CTXϕ, depends on the formation of a HJ by XerD catalysis, which is then resolved by XerC catalysis. The reaction nevertheless escapes the normal cellular control exerted by FtsK on XerD. In addition, we show that the same reaction promotes the excision of TLCϕ, along with any CTXϕ copy present between dif and its left attachment site, providing a plausible mechanism for how chrI CTXϕ copies can be eliminated, as occurred in the second wave of the current cholera pandemic.
与大多数细菌一样,霍乱弧菌的两条染色体chrI和chrII呈环状,由此产生的拓扑学问题通过两种酪氨酸重组酶XerC和XerD在每条染色体的特定位点dif处添加一个交叉来解决。该反应受细胞分裂蛋白FtsK的控制,FtsK通过XerD催化激活霍利迪连接体(HJ)中间体的形成,然后由XerC催化将其分解为产物。许多质粒和噬菌体分别利用Xer重组进行二聚体拆分和整合。在目前描述的所有情况下,它们都依赖于一种替代重组途径,其中XerC独立于FtsK催化HJ的形成。霍乱毒素噬菌体CTXϕ就是这种情况。在这里,我们表明,与之相反,TLCϕ(一种毒素相关的隐匿性卫星噬菌体,几乎总是在CTXϕ之前整合在chrI dif位点)的整合依赖于XerD催化形成HJ,然后由XerC催化将其分解。然而,该反应不受FtsK对XerD施加的正常细胞控制。此外,我们表明相同的反应促进了TLCϕ的切除,以及dif与其左附着位点之间存在的任何CTXϕ拷贝的切除,这为当前霍乱大流行第二波中chrI CTXϕ拷贝如何被消除提供了一种合理的机制。