Lease Richard A, Smith Dorie, McDonough Kathleen, Belfort Marlene
Wadsworth Center, New York State Department of Health, Center for Medical Sciences, Albany, New York, USA.
J Bacteriol. 2004 Sep;186(18):6179-85. doi: 10.1128/JB.186.18.6179-6185.2004.
DsrA RNA is a small (87-nucleotide) regulatory RNA of Escherichia coli that acts by RNA-RNA interactions to control translation and turnover of specific mRNAs. Two targets of DsrA regulation are RpoS, the stationary-phase and stress response sigma factor (sigmas), and H-NS, a histone-like nucleoid protein and global transcription repressor. Genes regulated globally by RpoS and H-NS include stress response proteins and virulence factors for pathogenic E. coli. Here, by using transcription profiling via DNA arrays, we have identified genes induced by DsrA. Steady-state levels of mRNAs from many genes increased with DsrA overproduction, including multiple acid resistance genes of E. coli. Quantitative primer extension analysis verified the induction of individual acid resistance genes in the hdeAB, gadAX, and gadBC operons. E. coli K-12 strains, as well as pathogenic E. coli O157:H7, exhibited compromised acid resistance in dsrA mutants. Conversely, overproduction of DsrA from a plasmid rendered the acid-sensitive dsrA mutant extremely acid resistant. Thus, DsrA RNA plays a regulatory role in acid resistance. Whether DsrA targets acid resistance genes directly by base pairing or indirectly via perturbation of RpoS and/or H-NS is not known, but in either event, our results suggest that DsrA RNA may enhance the virulence of pathogenic E. coli.
DsrA RNA是大肠杆菌中的一种小(87个核苷酸)调节性RNA,它通过RNA-RNA相互作用来控制特定mRNA的翻译和周转。DsrA调节的两个靶标是RpoS,即稳定期和应激反应σ因子(σ因子),以及H-NS,一种组蛋白样类核蛋白和全局转录抑制因子。由RpoS和H-NS全局调节的基因包括应激反应蛋白和致病性大肠杆菌的毒力因子。在这里,通过使用DNA阵列进行转录谱分析,我们鉴定了由DsrA诱导的基因。随着DsrA过量产生,许多基因的mRNA稳态水平增加,包括大肠杆菌的多个耐酸基因。定量引物延伸分析证实了hdeAB、gadAX和gadBC操纵子中单个耐酸基因的诱导。大肠杆菌K-12菌株以及致病性大肠杆菌O157:H7在dsrA突变体中表现出耐酸性受损。相反,从质粒过量产生DsrA使酸敏感的dsrA突变体具有极强的耐酸性。因此,DsrA RNA在耐酸性中起调节作用。DsrA是通过碱基配对直接靶向耐酸基因,还是通过干扰RpoS和/或H-NS间接靶向耐酸基因尚不清楚,但无论哪种情况,我们的结果都表明DsrA RNA可能增强致病性大肠杆菌的毒力。