AUTHOR=Deng Yiqin , Su Youlu , Liu Songlin , Guo Zhixun , Cheng Changhong , Ma Hongling , Wu Jinjun , Feng Juan , Chen Chang 

TITLE=Identification of a Novel Small RNA srvg23535 in Vibrio alginolyticus ZJ-T and Its Characterization With Phenotype MicroArray Technology

JOURNAL=Frontiers in Microbiology

VOLUME=9

YEAR=2018

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2018.02394

DOI=10.3389/fmicb.2018.02394

ISSN=1664-302X

ABSTRACT=<p>Small non-coding RNAs (sRNAs) are important modulators of gene expression and are involved in the pathogenesis and survival of prokaryotes. However, few studies have been conducted with <italic>Vibrio alginolyticus</italic>, which limits our ability to probe the regulation of virulence and environmental adaptation by sRNAs in this opportunistic pathogen. In this study, the sRNA candidate <italic>srvg23535</italic> was identified in <italic>V. alginolyticus</italic> ZJ-T. The precise transcript end, secondary structure, and sequence conservation were determined. A <italic>srvg23535</italic> null mutant was constructed and characterized by using Phenotype MicroArray (PM) technology. <italic>In silico</italic> target prediction was conducted by IntaRNA and TargetRNA2. Subsequently, a 107 nt transcript was validated with a sigma70 promoter at the 5′ end and a Rho-independent terminator at the 3′ end. The sRNA <italic>srvg23535</italic> had four stem-loop structures and was conserved among <italic>Vibrio harveyi, Vibrio parahaemolyticus</italic>, and <italic>Vibrio splendidus</italic>. Deletion of <italic>srvg23535</italic> in <italic>V. alginolyticus</italic> ZJ-T led to a weaker utilization of <sc>D</sc>-mannose, <sc>D</sc>-melibiose, lactulose, and inosine as carbon sources but stronger utilization of <sc>L</sc>-cysteine as nitrogen source. Moreover, the <italic>srvg2353</italic> mutant showed stronger resistance to osmotic stress but weaker resistance to pH stress. Additionally, a total of 22 common targets were identified and several were related to the observed phenotype of the mutant. This study indicated that the novel sRNA, <italic>srvg23535</italic>, is conserved and restricted to <italic>Vibrio</italic> spp., affecting the utilization of several carbon and nitrogen sources and the response to osmotic and pH stress. These results extend our understanding of sRNA regulation in <italic>V. alginolyticus</italic> and provide a significant resource for the further study of the precise target mRNAs of <italic>srvg23535</italic>, which may provide targets for antibacterial therapeutic or attenuated vaccines against <italic>Vibrio</italic> spp.</p>