RsmW, Pseudomonas aeruginosa small non-coding RsmA-binding RNA upregulated in biofilm versus planktonic growth conditions.

Autor: Miller CL; Dental and Craniofacial Trauma Research and Tissue Regeneration Directorate, Institute of Surgical Research, JBSA Fort Sam Houston, TX, USA., Romero M; School of Life Sciences, Centre for Biomolecular Sciences, University Park, University of Nottingham, Nottingham, NG7 2RD, UK., Karna SL; Dental and Craniofacial Trauma Research and Tissue Regeneration Directorate, Institute of Surgical Research, JBSA Fort Sam Houston, TX, USA., Chen T; The Forsyth Institute, Cambridge, Massachusetts, USA., Heeb S; School of Life Sciences, Centre for Biomolecular Sciences, University Park, University of Nottingham, Nottingham, NG7 2RD, UK., Leung KP; Dental and Craniofacial Trauma Research and Tissue Regeneration Directorate, Institute of Surgical Research, JBSA Fort Sam Houston, TX, USA. kai.p.leung.civ@mail.mil.
Jazyk: angličtina
Zdroj: BMC microbiology [BMC Microbiol] 2016 Jul 19; Vol. 16 (1), pp. 155. Date of Electronic Publication: 2016 Jul 19.
DOI: 10.1186/s12866-016-0771-y
Abstrakt: Background: Biofilm development, specifically the fundamentally adaptive switch from acute to chronic infection phenotypes, requires global regulators and small non-coding regulatory RNAs (sRNAs). This work utilized RNA-sequencing (RNA-seq) to detect sRNAs differentially expressed in Pseudomonas aeruginosa biofilm versus planktonic state.
Results: A computational algorithm was devised to detect and categorize sRNAs into 5 types: intergenic, intragenic, 5'-UTR, 3'-UTR, and antisense. Here we report a novel RsmY/RsmZ-type sRNA, termed RsmW, in P. aeruginosa up-transcribed in biofilm versus planktonic growth. RNA-Seq, 5'-RACE and Mfold predictions suggest RsmW has a secondary structure with 3 of 7 GGA motifs located on outer stem loops. Northern blot revealed two RsmW binding bands of 400 and 120 bases, suggesting RsmW is derived from the 3'-UTR of the upstream hypothetical gene, PA4570. RsmW expression is elevated in late stationary versus logarithmic growth phase in PB minimal media, at higher temperatures (37 °C versus 28 °C), and in both gacA and rhlR transposon mutants versus wild-type. RsmW specifically binds to RsmA protein in vitro and restores biofilm production and reduces swarming in an rsmY/rsmZ double mutant. PA4570 weakly resembles an RsmA/RsmN homolog having 49 % and 51 % similarity, and 16 % and 17 % identity to RsmA and RsmN amino acid sequences, respectively. PA4570 was unable to restore biofilm and swarming phenotypes in ΔrsmA deficient strains.
Conclusion: Collectively, our study reveals an interesting theme regarding another sRNA regulator of the Rsm system and further unravels the complexities regulating adaptive responses for Pseudomonas species.
Databáze: MEDLINE
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