Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire, USA
Department of Biological Sciences, Presidency University, Kolkata, India.
Infect Immun. 2018 Jan 22;86(2). doi: 10.1128/IAI.00635-17. Print 2018 Feb.
Expression of virulence factors in is regulated by a wide range of transcriptional regulators, including proteins and small RNAs (sRNAs), at the level of transcription and/or translation. The locus consists of three overlapping transcripts generated from three distinct promoters, all containing the open reading frame (ORF). The 5' untranslated regions (UTRs) of these transcripts contain three separate regions ∼711, 409, and 146 nucleotides (nt) upstream of the translation start, the functions of which remain unknown. Recent transcriptome-sequencing (RNA-Seq) analysis and subsequent characterization indicated that two sRNAs, teg49 and teg48, are processed and likely produced from the P3 and P1 transcripts of the locus, respectively. In this report, we utilized a variety of promoter mutants and and mutants to ascertain the contributions of these factors to the generation of teg49. We also defined the transcriptional regulon of teg49, including virulence genes not regulated by SarA. Phenotypically, teg49 did not impact biofilm formation or affect overall SarA expression significantly. Comparative analyses of RNA-Seq data between the wild-type, teg49 mutant, and mutant strains indicated that ∼133 genes are significantly upregulated while 97 are downregulated in a teg49 deletion mutant in a -independent manner. An abscess model of skin infection indicated that the teg49 mutant exhibited a reduced bacterial load compared to the wild-type Overall, these results suggest that teg49 sRNA has a regulatory role in target gene regulation independent of SarA. The exact mechanism of this regulation is yet to be dissected.
毒力因子的表达在 水平上受到广泛的转录调控因子的调控,包括蛋白质和小 RNA(sRNA),在转录和/或翻译水平上。 基因座由三个重叠的转录本组成,这些转录本由三个不同的启动子产生,均包含 开放阅读框(ORF)。这些转录本的 5'非翻译区(UTR)包含三个独立的区域,在 翻译起始点上游约 711、409 和 146 个核苷酸(nt),其功能未知。最近的转录组测序(RNA-Seq)分析和随后的表征表明,两个 sRNA,teg49 和 teg48,分别从 基因座的 P3 和 P1 转录物加工而来。在本报告中,我们利用各种 启动子突变体和 和 突变体来确定这些因素对 teg49 产生的贡献。我们还定义了 teg49 的转录调控因子,包括不受 SarA 调控的毒力基因。表型上,teg49 对生物膜形成没有影响,也没有显著影响 SarA 的整体表达。野生型、teg49 突变体和 突变体菌株之间的 RNA-Seq 数据比较分析表明,在 teg49 缺失突变体中,约有 133 个基因以 -独立的方式显著上调,97 个基因下调。皮肤感染脓肿模型表明,与野生型 相比,teg49 突变体的细菌负荷降低。总的来说,这些结果表明 teg49 sRNA 在不依赖 SarA 的情况下在靶基因调控中具有调节作用。这种调节的确切机制尚未被剖析。