Discovering genetic determinants for cell-to-cell adhesion in two prevalent conjugative lactococcal plasmids

IF 4.8 Q1 MICROBIOLOGY Current Research in Microbial Sciences Pub Date : 2024-01-01 DOI:10.1016/j.crmicr.2024.100239
Guillermo Ortiz Charneco , Philip Kelleher , Andrius Buivydas , Paul P. de Waal , Irma M.H. van Rijswijck , Noël N.M.E. van Peij , Christian Cambillau , Jennifer Mahony , Douwe Van Sinderen
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Abstract

Plasmids pNP40 and pUC11B encode two prevalent yet divergent conjugation systems, which have been characterized in detail recently. Here, we report the elucidation of the putative adhesins of the pNP40 and pUC11B conjugation systems, encoded by traAd and trsAd, respectively. Despite their significant sequence divergence, TraAd and TrsAd represent the most conserved component between the pNP40- and the pUC11B-encoded conjugation systems and share similar peptidoglycan-hydrolase domains. Protein structure prediction using AlphaFold2 highlighted the structural similarities between their predicted domains, as well as the potential homo-dimeric state of both proteins. Expression of the putative surface adhesins resulted in a cell clumping phenotype not only among cells expressing these surface adhesins but also between adhesin-expressing and non-producing cells. Furthermore, mutant derivatives of plasmids pNP40 or pUC11B carrying a mutation in traAd or trsAd, respectively, were shown to act as efficient donors provided the corresponding recipient expresses either traAd or trsAd, thus demonstrating in trans reciprocal complementarity of these proteins in conjugation systems.

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在两种流行的共轭乳球菌质粒中发现细胞间粘附的遗传决定因素
质粒 pNP40 和 pUC11B 编码两种普遍存在但又存在差异的连接系统,最近对这两种系统进行了详细表征。在这里,我们报告了分别由 traAd 和 trsAd 编码的 pNP40 和 pUC11B 连接系统的假定粘附素的阐明。尽管它们的序列差异很大,但 TraAd 和 TrsAd 代表了 pNP40 和 pUC11B 连接系统之间最保守的成分,并且具有相似的肽聚糖水解酶结构域。利用 AlphaFold2 进行的蛋白质结构预测突显了其预测结构域之间的结构相似性,以及这两种蛋白质潜在的同源二聚体状态。表达推测的表面粘附蛋白不仅会在表达这些表面粘附蛋白的细胞之间,而且会在表达粘附蛋白的细胞和不表达粘附蛋白的细胞之间产生细胞聚集表型。此外,分别携带 traAd 或 trsAd 突变的质粒 pNP40 或 pUC11B 的突变衍生物在相应的受体表达 traAd 或 trsAd 的情况下可作为有效的供体,从而证明了这些蛋白在共轭体系中的反式互补性。
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来源期刊
Current Research in Microbial Sciences
Current Research in Microbial Sciences Immunology and Microbiology-Immunology and Microbiology (miscellaneous)
CiteScore
7.90
自引率
0.00%
发文量
81
审稿时长
66 days
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