Widespread targeting of nascent transcripts by RsmA in Pseudomonas aeruginosa

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 117; no. 19; pp. 10520 - 10529
• Main Authors: Gebhardt, Michael J., Kambara, Tracy K., Ramsey, Kathryn M., Dove, Simon L.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 12.05.2020
• Subjects: Biofilms; Biological Sciences; Chromatin; Combinatorial analysis; Deoxyribonucleic acid; DNA; DNA sequencing; DNA-directed RNA polymerase; Immunoprecipitation; Opportunist infection; Post-transcription; Proteins; Pseudomonas aeruginosa; Regulators; Ribonucleic acid; Rifampin; RNA; RNA polymerase; RNA-binding protein; Virulence; posttranscriptional regulator; Hfq; RNA binding proteins
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1917587117

In the opportunistic pathogen Pseudomonas aeruginosa, 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 P. aeruginosa. 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 P. aeruginosa, 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.