Genome-wide screen and functional analysis in Xanthomonas reveal a large number of mRNA-derived sRNAs, including the novel RsmA-sequester RsmU.
Genome-wide screen and functional analysis in Xanthomonas reveal a large number of mRNA-derived sRNAs, including the novel RsmA-sequester RsmU.
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黄单胞菌基因组宽屏和功能分析揭示了大量 mRNA 衍生的 sRNA，包括新型 RsmA 隔离器 RsmU
DOI：
10.1111/mpp.12997
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发表时间：
2020-12
影响因子：
4.9
通讯作者：
Tang JL
中科院分区：
文献类型：
--
作者：
Tang DJ;Chen XL;Jia Y;Liang YW;He YP;Lu TT;Zhu CR;Han B;An SQ;Tang JL
尽管已知细菌小非编码rna （sRNAs）在各种细胞过程中发挥关键作用，包括发病机制，但在许多生物体中，这些sRNAs的身份和作用仍然知之甚少。在这里，我们对油菜黄单胞菌pv的srna进行了全基因组筛选和功能分析。campestris （Xcc）是一种重要的植物病原体。对在毒力基因诱导条件下生长的野生型菌株分离的50-500 - nt RNA片段进行测序，共鉴定出612个sRNA候选（src）。大多数（82%）SRCs来源于mRNA，而不是特定的sRNA基因。采用northern印迹法对121例有代表性的src进行分析；检测到117个src，支持了612个src中绝大多数确实是srna的论点。对过表达不同候选菌株的表型分析表明，一种特定的sRNA RsmU在Xcc中起着毒力、超敏反应和细胞运动的负调节作用。尽管序列分析显示RsmU不是已知的拮抗RsmA的sRNAs家族的成员，但体外电泳迁移率转移试验和体内共免疫沉淀分析表明，RsmU与全球转录后调节因子RsmA相互作用。Northern blotting分析表明，RsmU有两个同工型，它们是从XC1332 mRNA的3 '‐未翻译区域加工而成，预计会编码ComEA， ComEA是细菌摄取DNA所需的一种质周蛋白。这项工作揭示了细菌中意想不到的主要sRNA生物发生策略，以及sRNA介导的Xcc毒力调节的隐藏层。来自XC1332 mRNA 3′‐UTR的RsmA隔离非编码RNA RsmU负调控油菜黄单胞菌的毒力、超敏反应和细胞运动。
Although bacterial small noncoding RNAs (sRNAs) are known to play a critical role in various cellular processes, including pathogenesis, the identity and action of such sRNAs are still poorly understood in many organisms. Here we have performed a genome‐wide screen and functional analysis of the sRNAs in Xanthomonas campestris pv. campestris (Xcc), an important phytopathogen. The 50–500‐nt RNA fragments isolated from the wild‐type strain grown in a virulence gene‐inducing condition were sequenced and a total of 612 sRNA candidates (SRCs) were identified. The majority (82%) of the SRCs were derived from mRNA, rather than specific sRNA genes. A representative panel of 121 SRCs were analysed by northern blotting; 117 SRCs were detected, supporting the contention that the overwhelming majority of the 612 SRCs identified are indeed sRNAs. Phenotypic analysis of strains overexpressing different candidates showed that a particular sRNA, RsmU, acts as a negative regulator of virulence, the hypersensitive response, and cell motility in Xcc. In vitro electrophoretic mobility shift assay and in vivo coimmunoprecipitation analyses indicated that RsmU interacted with the global posttranscriptional regulator RsmA, although sequence analysis displayed that RsmU is not a member of the sRNAs families known to antagonize RsmA. Northern blotting analyses demonstrated that RsmU has two isoforms that are processed from the 3′‐untranslated region of the mRNA of XC1332 predicted to encode ComEA, a periplasmic protein required for DNA uptake in bacteria. This work uncovers an unexpected major sRNA biogenesis strategy in bacteria and a hidden layer of sRNA‐mediated virulence regulation in Xcc. The RsmA‐sequestering noncoding RNA RsmU derived from the 3′‐UTR of XC1332 mRNA negatively regulates virulence, the hypersensitive response, and cell motility in Xanthomonas campestris.
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影响因子：
11.3
作者：
An SQ;Potnis N;Dow M;Vorhölter FJ;He YQ;Becker A;Teper D;Li Y;Wang N;Bleris L;Tang JL
通讯作者：
Tang JL
影响因子：
5.7
作者：
Drecktrah D;Hall LS;Rescheneder P;Lybecker M;Samuels DS
通讯作者：
Samuels DS
影响因子：
14.9
作者：
Kalvari I;Argasinska J;Quinones-Olvera N;Nawrocki EP;Rivas E;Eddy SR;Bateman A;Finn RD;Petrov AI
通讯作者：
Petrov AI
影响因子：
2.1
作者：
Ishikawa, J;Hotta, K
通讯作者：
Hotta, K
DOI：
10.1038/nrm4085
发表时间：
2015-12
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
Borges F;Martienssen RA
通讯作者：
Martienssen RA