Division of Infectious Diseases, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115.
Division of Infectious Diseases, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115
Proc Natl Acad Sci U S A. 2020 May 12;117(19):10520-10529. doi: 10.1073/pnas.1917587117. Epub 2020 Apr 24.
In the opportunistic pathogen , RsmA is an RNA-binding protein that plays critical roles in the control of virulence, interbacterial interactions, and biofilm formation. Although RsmA is thought to exert its regulatory effects by binding full-length transcripts, the extent to which RsmA binds nascent transcripts has not been addressed. Moreover, which transcripts are direct targets of this key posttranscriptional regulator is largely unknown. Using chromatin immunoprecipitation coupled with high-throughput DNA sequencing, with cells grown in the presence and absence of the RNA polymerase inhibitor rifampicin, we identify hundreds of nascent transcripts that RsmA associates with in We also find that the RNA chaperone Hfq targets a subset of those nascent transcripts that RsmA associates with and that the two RNA-binding proteins can exert regulatory effects on common targets. Our findings establish that RsmA associates with many transcripts as they are being synthesized in , identify the transcripts targeted by RsmA, and suggest that RsmA and Hfq may act in a combinatorial fashion on certain transcripts. The binding of posttranscriptional regulators to nascent transcripts may be commonplace in bacteria where distinct regulators can function alone or in concert to achieve control over the translation of transcripts as soon as they emerge from RNA polymerase.
在机会致病菌中,RsmA 是一种 RNA 结合蛋白,在控制毒力、细菌间相互作用和生物膜形成方面发挥着关键作用。尽管 RsmA 被认为通过结合全长转录本发挥其调节作用,但 RsmA 结合新生转录本的程度尚未得到解决。此外,该关键转录后调节因子的直接靶标在很大程度上尚不清楚。使用染色质免疫沉淀结合高通量 DNA 测序,在存在和不存在 RNA 聚合酶抑制剂利福平的情况下培养细胞,我们鉴定了数百个 RsmA 与之结合的新生转录本。我们还发现 RNA 伴侣 Hfq 靶向 RsmA 与之结合的部分新生转录本,并且这两个 RNA 结合蛋白可以对共同的靶标发挥调节作用。我们的发现确立了 RsmA 在合成过程中与许多转录本结合,鉴定了 RsmA 的靶标转录本,并表明 RsmA 和 Hfq 可能以组合方式作用于某些转录本。在细菌中,转录后调节因子与新生转录本的结合可能很常见,其中不同的调节因子可以单独或协同作用,以控制从 RNA 聚合酶中出现的转录本的翻译。