Concerted spatio-temporal dynamics of imported DNA and ComE DNA uptake protein during gonococcal transformation.
Concerted spatio-temporal dynamics of imported DNA and ComE DNA uptake protein during gonococcal transformation.
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DOI：
10.1371/journal.ppat.1004043
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发表时间：
2014-04
期刊：
影响因子：
6.7
通讯作者：
Maier B
中科院分区：
文献类型：
--
作者：
Gangel H;Hepp C;Müller S;Oldewurtel ER;Aas FE;Koomey M;Maier B
转化能力在细菌物种中广泛存在。在革兰氏阴性菌的情况下，转化的关键步骤是通过外膜输入DNA。虽然已知有多种因素影响DNA转运，但对DNA进口的动态了解甚少。在这里，我们描述了DNA进入淋病奈瑟菌周质的时空动态。DNA被导入到细胞轮廓周围随机位置的周质中。随后，它被招募到双球菌的隔膜，其时间尺度随着DNA长度的增加而增加。我们用荧光DNA发现，用∼40kbpDNA在几分钟内就能使周质饱和。DNA结合蛋白以基因剂量依赖的方式定量地控制周质的运载能力。使用荧光标记的衍生蛋白可以看到，在没有外源DNA的情况下，Come均匀地分布在周质中。在加入外源DNA后，Come被重新定位，形成与进口DNA共定位的离散灶。我们的结论是，周质可以作为进口DNA的相当大的储存库，控制着潜在地储存的DNA的量，以便通过内膜运输。细菌转化是外源DNA的导入和遗传整合。因此，它被认为是一种主要的进化力量。关键的一步是通过外膜输入DNA。在这里，我们描述了DNA在输入和停留在革兰氏阴性病原体淋球菌周质中的时空动态。我们发现，周质可以作为进口DNA的储存库，通过从环境中进口DNA，这些DNA可以在5分钟内填满。进口DNA的数量大致相当于一个噬菌体基因组的大小。在没有胞外DNA的情况下，周质DNA结合蛋白COME均匀地分布在周质中。当将外部DNA添加到合格的淋球菌中时，它会迅速重新定位到进口DNA上。由于控制着周质的承载能力，我们认为它可能会凝聚DNA，从而将DNA摄取与其紧凑联系在一起。虽然通过外膜输入的DNA局限于细胞轮廓周围，但大部分输入的DNA重新定位到双球菌中心的隔膜。我们的发现有力地支持了这样一种观点，即周围质团的DNA不依赖于通过内膜的运输。
Competence for transformation is widespread among bacterial species. In the case of Gram-negative systems, a key step to transformation is the import of DNA across the outer membrane. Although multiple factors are known to affect DNA transport, little is known about the dynamics of DNA import. Here, we characterized the spatio-temporal dynamics of DNA import into the periplasm of Neisseria gonorrhoeae. DNA was imported into the periplasm at random locations around the cell contour. Subsequently, it was recruited at the septum of diplococci at a time scale that increased with DNA length. We found using fluorescent DNA that the periplasm was saturable within minutes with ∼40 kbp DNA. The DNA-binding protein ComE quantitatively governed the carrying capacity of the periplasm in a gene-dosage-dependent fashion. As seen using a fluorescent-tagged derivative protein, ComE was homogeneously distributed in the periplasm in the absence of external DNA. Upon addition of external DNA, ComE was relocalized to form discrete foci colocalized with imported DNA. We conclude that the periplasm can act as a considerable reservoir for imported DNA with ComE governing the amount of DNA stored potentially for transport through the inner membrane. Bacterial transformation is the import and inheritable integration of external DNA. As such, it is believed to be a major evolutionary force. A key step is the import of DNA through the outer membrane. Here, we have characterized the spatio-temporal dynamics of DNA during import and residence in the periplasm of the Gram-negative pathogen Neisseria gonorrhoeae. We found that the periplasm can serve as a reservoir for imported DNA that can fill within five minutes by importing DNA from the environment. The amount of imported DNA roughly corresponds to the size of a phage genome. The periplasmic DNA-binding protein ComE is homogeneously distributed in the periplasm in the absence of extracellular DNA. It relocates rapidly to imported DNA when external DNA is added to competent gonococci. As ComE governs the carrying capacity of the periplasm, we propose that it might condense DNA, thus linking DNA uptake to its compaction. Although the import through the outer membrane was localized all around the cell contour, the major part of the imported DNA relocated to the septum at the center of diplococci. Our findings strongly support the idea that the periplasm masses DNA independently of transport through the inner membrane.
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影响因子：
4.5
作者：
Attaiech L;Olivier A;Mortier-Barrière I;Soulet AL;Granadel C;Martin B;Polard P;Claverys JP
通讯作者：
Claverys JP
DOI：
10.3109/10409238.2012.729562
发表时间：
2012-11
影响因子：
6.5
作者：
Kidane D;Ayora S;Sweasy JB;Graumann PL;Alonso JC
通讯作者：
Alonso JC
DOI：
10.1073/pnas.1218832110
发表时间：
2013-02-19
影响因子：
11.1
作者：
Cehovin, Ana;Simpson, Peter J.;Pelicic, Vladimir
通讯作者：
Pelicic, Vladimir
影响因子：
7.7
作者：
Allemand, Jean-Francois;Maier, Berenike;Smith, Douglas E.
通讯作者：
Smith, Douglas E.
影响因子：
8.6
作者：
Holz, Claudia;Opitz, Dirk;Maier, Berenike
通讯作者：
Maier, Berenike