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Identification of ComW as a new component in the regulation of genetic transformation in Streptococcus pneumoniae

机译:鉴定ComW作为肺炎链球菌遗传转化调控的新成分

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摘要

Regulation of competence for genetic transformation in Streptococcus pneumoniae depends on a quorum-sensing system, genes involved in DNA uptake and recombination and a link between these two gene sets. The alternative sigma factor ComX provides this link. ComE, the response regulator of the quorum-sensing system, is required for expression of ComX and other early genes. However, an unknown ComE-dependent regulator is also required for competence when comX is expressed under control of the raffinose-responsive promoter of the aga operon. The gene comW (SP0018) is required for a high level of competence and is regulated by the quorum-sensing system, but its function is unknown. To explore its role further, comW was cloned into the multicopy plasmid pMSP3535, under the control of a nisin-inducible promoter (P-N), and transformed into pneumococcal strains containing a raffinose-inducible comX gene (P-R::comX). Further introduction of a comE deletion blocked the endogenous CSP signal transduction pathway. In the resulting strain, competence was independent of CSP but depended on treatment with both nisin and raffinose, showing that coexpression of comW and comX complemented the comE deficiency. ComX protein accumulation and expression of a late competence gene in the above strain support the conclusion that ComW is a new positive factor involved in competence regulation.
机译:肺炎链球菌遗传转化能力的调节取决于群体感应系统,参与DNA吸收和重组的基因以及这两个基因组之间的联系。替代的sigma factor ComX提供了此链接。 ComE是群体感应系统的响应调节器,对于ComX和其他早期基因的表达是必需的。但是,当在aga操纵子的棉子糖反应性启动子的控制下表达comX时,也需要未知的ComE依赖性调节剂来发挥作用。 comW(SP0018)基因是高水平能力所必需的,并由群体感应系统调节,但功能未知。为了进一步探讨其作用,在乳链菌肽诱导型启动子(P-N)的控制下,将comW克隆到多拷贝质粒pMSP3535中,并转化为含有棉子糖诱导型comX基因(P-R :: comX)的肺炎球菌菌株。进一步引入comE缺失可阻断内源性CSP信号转导途径。在产生的菌株中,能力不依赖于CSP,但依赖于乳链菌肽和棉子糖的治疗,这表明comW和comX的共表达弥补了comE缺陷。在上述菌株中,ComX蛋白的积累和后期能力基因的表达支持以下结论:ComW是参与能力调节的新的积极因素。

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