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Exploration of DNA processing features unravels novel properties of ICE conjugation in Gram-positive bacteria
Nucleic Acids Research ( IF 14.9 ) Pub Date : 2022-07-18 , DOI: 10.1093/nar/gkac607
Haifa Laroussi 1 , Yanis Aoudache 1 , Emilie Robert 1 , Virginie Libante 1 , Louise Thiriet 1 , Dominique Mias-Lucquin 2 , Badreddine Douzi 1 , Yvonne Roussel 1 , Isaure Chauvot de Beauchêne 2 , Nicolas Soler 1 , Nathalie Leblond-Bourget 1
Affiliation  

Integrative and conjugative elements (ICEs) are important drivers of horizontal gene transfer in prokaryotes. They are responsible for antimicrobial resistance spread, a major current health concern. ICEs are initially processed by relaxases that recognize the binding site of oriT sequence and nick at a conserved nic site. The ICESt3/Tn916/ICEBs1 superfamily, which is widespread among Firmicutes, encodes uncanonical relaxases belonging to a recently identified family called MOBT. This family is related to the rolling circle replication initiators of the Rep_trans family. The nic site of these MOBT relaxases is conserved but their DNA binding site is still unknown. Here, we identified the bind site of RelSt3, the MOBT relaxase from ICESt3. Unexpectedly, we found this bind site distantly located from the nic site. We revealed that the binding of the RelSt3 N-terminal HTH domain is required for efficient nicking activity. We also deciphered the role of RelSt3 in the initial and final stages of DNA processing during conjugation. Especially, we demonstrated a strand transfer activity, and the formation of covalent DNA-relaxase intermediate for a MOBT relaxase.

中文翻译:

DNA加工特征的探索揭示了革兰氏阳性菌中ICE结合的新特性

整合和共轭元件 (ICE) 是原核生物水平基因转移的重要驱动因素。它们负责抗菌素耐药性的传播,这是当前的主要健康问题。ICE 最初由识别 oriT 序列的结合位点和在保守 nic 位点产生切口的松弛酶处理。ICESt3/Tn916/ICEBs1 超家族在厚壁菌门中广泛存在,它编码属于最近发现的一个名为 MOBT 的家族的非典型松弛酶。该族与 Rep_trans 族的滚环复制发起者有关。这些 MOBT 松弛酶的 nic 位点是保守的,但它们的 DNA 结合位点仍然未知。在这里,我们确定了来自 ICESt3 的 MOBT 松弛酶 RelSt3 的结合位点。出乎意料的是,我们发现这个绑定站点远离 nic 站点。我们揭示了 RelSt3 N 末端 HTH 结构域的结合是有效切口活动所必需的。我们还破译了 RelSt3 在结合过程中 DNA 加工的初始和最后阶段的作用。特别是,我们展示了链转移活性,以及​​ MOBT 松弛酶的共价 DNA-松弛酶中间体的形成。
更新日期:2022-07-18
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