Irazoki Oihane, Aranda Jesús, Zimmermann Timo, Campoy Susana, Barbé Jordi
Department de Genètica i de Microbiologia, Universitat Autònoma de Barcelona Cerdanyola del Vallès, Spain.
Advanced Light Microscopy Unit, Center for Genomic Regulation Barcelona, Spain.
Front Microbiol. 2016 Oct 6;7:1560. doi: 10.3389/fmicb.2016.01560. eCollection 2016.
In addition to its role in DNA damage repair and recombination, the RecA protein, through its interaction with CheW, is involved in swarming motility, a form of flagella-dependent movement across surfaces. In order to better understand how SOS response modulates swarming, in this work the location of RecA and CheW proteins within the swarming cells has been studied by using super-resolution microscopy. Further, and after docking studies, the specific RecA and CheW regions associated with the RecA-CheW interaction have also been confirmed by site-directed mutagenesis and immunoprecipitation techniques. Our results point out that the CheW distribution changes, from the cell poles to foci distributed in a helical pattern along the cell axis when SOS response is activated or RecA protein is overexpressed. In this situation, the CheW presents the same subcellular location as that of RecA, pointing out that the previously described RecA storage structures may be modulators of swarming motility. Data reported herein not only confirmed that the RecA-CheW pair is essential for swarming motility but it is directly involved in the CheW distribution change associated to SOS response activation. A model explaining not only the mechanism by which DNA damage modulates swarming but also how both the lack and the excess of RecA protein impair this motility is proposed.
RecA蛋白除了在DNA损伤修复和重组中发挥作用外,还通过与CheW相互作用参与群体游动,这是一种依赖鞭毛在表面移动的形式。为了更好地理解SOS反应如何调节群体游动,在这项工作中,通过使用超分辨率显微镜研究了群体游动细胞内RecA和CheW蛋白的定位。此外,在对接研究之后,还通过定点诱变和免疫沉淀技术证实了与RecA-CheW相互作用相关的特定RecA和CheW区域。我们的结果指出,当SOS反应被激活或RecA蛋白过表达时,CheW的分布会发生变化,从细胞极变为沿细胞轴呈螺旋模式分布的焦点。在这种情况下,CheW呈现出与RecA相同的亚细胞定位,表明先前描述的RecA储存结构可能是群体游动的调节因子。本文报道的数据不仅证实了RecA-CheW对对于群体游动至关重要,而且它直接参与了与SOS反应激活相关的CheW分布变化。提出了一个模型,该模型不仅解释了DNA损伤调节群体游动的机制,还解释了RecA蛋白的缺乏和过量如何损害这种游动。