Gruber Charley C, Sperandio Vanessa
Department of Microbiology and Department of Biochemistry, UT Southwestern Medical Center, Dallas, Texas, USA.
Department of Microbiology and Department of Biochemistry, UT Southwestern Medical Center, Dallas, Texas, USA
Infect Immun. 2015 Apr;83(4):1286-95. doi: 10.1128/IAI.02918-14. Epub 2015 Jan 20.
Enterohemorrhagic Escherichia coli (EHEC) is a significant human pathogen and is the cause of bloody diarrhea and hemolytic-uremic syndrome. The virulence repertoire of EHEC includes the genes within the locus of enterocyte effacement (LEE) that are largely organized in five operons, LEE1 to LEE5, which encode a type III secretion system, several effectors, chaperones, and regulatory proteins. In addition, EHEC also encodes several non-LEE-encoded effectors and fimbrial operons. The virulence genes of this pathogen are under a large amount of posttranscriptional regulation. The small RNAs (sRNAs) GlmY and GlmZ activate the translation of glucosamine synthase (GlmS) in E. coli K-12, and in EHEC they destabilize the 3' fragments of the LEE4 and LEE5 operons and promote translation of the non-LEE-encoded effector EspFu. We investigated the global changes of EHEC gene expression governed by GlmY and GlmZ using RNA sequencing and gene arrays. This study extends the known effects of GlmY and GlmZ regulation to show that they promote expression of the curli adhesin, repress the expression of tryptophan metabolism genes, and promote the expression of acid resistance genes and the non-LEE-encoded effector NleA. In addition, seven novel EHEC-specific sRNAs were identified using RNA sequencing, and three of them--sRNA56, sRNA103, and sRNA350--were shown to regulate urease, fimbria, and the LEE, respectively. These findings expand the knowledge of posttranscriptional regulation in EHEC.
肠出血性大肠杆菌(EHEC)是一种重要的人类病原体,可导致血性腹泻和溶血尿毒综合征。EHEC的毒力基因库包括肠细胞脱落位点(LEE)内的基因,这些基因主要组织成五个操纵子,即LEE1至LEE5,它们编码III型分泌系统、几种效应蛋白、分子伴侣和调节蛋白。此外,EHEC还编码几种非LEE编码的效应蛋白和菌毛操纵子。该病原体的毒力基因受到大量转录后调控。小RNA(sRNA)GlmY和GlmZ可激活大肠杆菌K-12中氨基葡萄糖合酶(GlmS)的翻译,而在EHEC中,它们会使LEE4和LEE5操纵子的3'片段不稳定,并促进非LEE编码效应蛋白EspFu的翻译。我们使用RNA测序和基因芯片研究了由GlmY和GlmZ调控的EHEC基因表达的全局变化。这项研究扩展了已知的GlmY和GlmZ调控的作用,表明它们促进卷曲菌毛黏附素的表达,抑制色氨酸代谢基因的表达,并促进耐酸基因和非LEE编码效应蛋白NleA的表达。此外,通过RNA测序鉴定出了七种新的EHEC特异性sRNA,其中三种——sRNA56、sRNA103和sRNA350——分别被证明可调控脲酶、菌毛和LEE。这些发现扩展了对EHEC转录后调控的认识。