Assalkhou Reza, Balasingham Seetha, Collins Richard F, Frye Stephan A, Davidsen Tonje, Benam Afsaneh V, Bjørås Magnar, Derrick Jeremy P, Tønjum Tone
Centre for Molecular Biology and Neuroscience and Institute of Microbiology, Rikshospitalet-Radiumhospitalet Medical Centre, Oslo, Norway.
Centre for Molecular Biology and Neuroscience and Institute of Microbiology, University of Oslo, Oslo, Norway.
Microbiology (Reading). 2007 May;153(Pt 5):1593-1603. doi: 10.1099/mic.0.2006/004200-0.
Neisseria meningitidis is naturally competent for transformation throughout its growth cycle. Transformation in neisserial species is coupled to the expression of type IV pili, which are present on the cell surface as bundled filamentous appendages, and are assembled, extruded and retracted by the pilus biogenesis components. During the initial phase of the transformation process, binding and uptake of DNA takes place with entry through a presumed outer-membrane channel into the periplasm. This study showed that DNA associates only weakly with purified pili, but binds significantly to the PilQ complex isolated directly from meningococcal membranes. By assessing the DNA-binding activity of the native complex PilQ, as well as recombinant truncated PilQ monomers, it was shown that the N-terminal region of PilQ is involved in the interaction with DNA. It was evident that the binding of ssDNA to PilQ had a higher affinity than the binding of dsDNA. The binding of DNA to PilQ did not, however, depend on the presence of the neisserial DNA-uptake sequence. It is suggested that transforming DNA is introduced into the cell through the outer-membrane channel formed by the PilQ complex, and that DNA uptake occurs by non-specific introduction of DNA coupled to pilus retraction, followed by presentation to DNA-binding component(s), including PilQ.
脑膜炎奈瑟菌在其整个生长周期中都具有自然转化能力。奈瑟菌属中的转化与IV型菌毛的表达相关联,IV型菌毛以成束的丝状附属物形式存在于细胞表面,并由菌毛生物发生组件进行组装、挤出和回缩。在转化过程的初始阶段,DNA的结合和摄取通过一个推测的外膜通道进入周质而发生。这项研究表明,DNA与纯化的菌毛结合较弱,但与直接从脑膜炎球菌膜中分离出的PilQ复合物有显著结合。通过评估天然复合物PilQ以及重组截短的PilQ单体的DNA结合活性,结果表明PilQ的N端区域参与了与DNA的相互作用。很明显,单链DNA与PilQ的结合亲和力高于双链DNA。然而,DNA与PilQ的结合并不依赖于奈瑟菌DNA摄取序列的存在。有人提出,转化DNA是通过由PilQ复合物形成的外膜通道引入细胞的,并且DNA摄取是通过与菌毛回缩相关的非特异性DNA引入,随后呈现给包括PilQ在内的DNA结合成分。