Journal
BMC MICROBIOLOGY
Volume 16, Issue -, Pages -Publisher
BMC
DOI: 10.1186/s12866-016-0771-y
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Funding
- Naval Medical Research Center's Advanced Medical Development Program [MIPR N3239815MHX040]
- US Army Medical Research and Materiel Command, Dental and Craniofacial Trauma Research and Tissue Regeneration Directorate
- fellowship Apoio a formacion posdoutoral do PLAN I2C da Xunta de Galicia
- [NIH P30 DK089507]
- Engineering and Physical Sciences Research Council [EP/N006615/1] Funding Source: researchfish
- EPSRC [EP/N006615/1] Funding Source: UKRI
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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 degrees C versus 28 degrees 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 Delta rsmA 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.
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