细菌中的小RNA

小RNA（sRNA）或非编码RNA（ncRNA）在原核生物和真核生物中广泛分布。迄今,在各种细菌中共发现超过150种sRNA,在大肠杆菌中发现了约80种sRNA。sRNA通过与靶mRNA配对而发生作用,导致mRNA翻译和稳定性的变化;sRNA的功能涉及从结构调节到催化作用,影响生物体内各种各样的加工过程,一个单独的sRNA就能调控大量的基因并对细胞生理产生深远影响。目前,对sRNA的研究主要采用生物信息学预测结合分子生物学实验的方法。     

小RNA对胞内菌生长、毒力及铁水平的调节作用

细菌小RNA(small RNA,sRNA)是一类长度为50~500个碱基,具有调控转录、翻译和mRNA稳定性的非编码调节性RNA。随着越来越多的sRNA被鉴定,部分细菌的sRNA功能已逐步阐明,主要参与调控细菌的基因表达、增殖、毒力及对环境的应激反应等生物学过程。本文就胞内菌(如沙门菌、李斯特菌、嗜肺军团菌等)sRNA对其自身在宿主细胞内的生长、毒力和铁水平的调控作用进行综述。     

细菌非编码小RNA的筛选及鉴定方法

细菌非编码小RNA（smallnon．codingRNAs,sRNAs）是一类长度为50～500nt、不编码蛋白质的功能RNA,在应对胁迫、毒力产生和新陈代谢等生命过程中起重要的调控作用。其主要通过碱基配对与靶mRNA发生作用,导致mRNA翻译和稳定性改变,从而在转录后水平调节基因的表达,最终影响细菌各种生命活动。近年来,利用生物信息学和分子生物学技术,已在细菌中筛选并鉴定得到了几百个sRNA。该文对细菌sRNA的筛选和鉴定方法作一简要综述。     

细菌sRNA数据库

细菌sRNA是一类长度在50～500 nt的调控小RNA（small regulatory RNA）,主要通过与靶标mRNA或靶标蛋白质结合发挥多种生物学功能。目前,随着生物信息学与高通量测序的应用,发现了越来越多的细菌sRNA,开发了多个相关数据库。为了sRNA工作者系统了解与应用这些数据,本文拟对包含细菌sRNA的综合数据库和细菌sRNA专业数据库作一概述,并对sRNA数据库的未来发展进行展望。     

细菌非编码小RNA功能及作用机制研究进展

生物体中除了编码蛋白质的mRNA外,还存在多种具有重要调控功能的非编码RNA。细菌中长度50~500 nt的非编码RNA通常定义为sRNA。sRNA在细菌的整个生命活动中发挥着极为广泛的作用,在感受环境压力、基因表达、细胞周期乃至个体发育等过程中均具有重要的调控作用。sRNA的功能学和调控机制的研究已成为当今细菌学研究的热点。本研究就细菌中的sRNA的特征,在细菌中的作用和作用机制进行文献综述。     

细菌小RNA的研究

小RNA(smallRNA,sRNA)在基因表达调控和生长发育等方面发挥着重要作用。细菌sRNA多通过与靶mRNA配对,转录后水平影响目的mRNA翻译或(和)稳定性,对基因的表达进行调节,以影响细胞的多种生理功能。本文从细菌sRNA与真核生物微RNA(microRNA,miRNA)的比较,sRNA的分类,sRNA分子伴侣Hfq及sRNA鉴别方法等方面综述了sRNA的研究进展,指出目前sRNA研究仍然存在的问题。原核生物中sRNA的大量发现和深入研究,有可能使人们对生物进化和生命的发展过程有更为深入的认识与了解。     

细菌sRNA数据库北大核心CSCD

细菌sRNA是一类长度在50~500 nt的调控小RNA(small regulatory RNA),主要通过与靶标mRNA或靶标蛋白质结合发挥多种生物学功能。目前,随着生物信息学与高通量测序的应用,发现了越来越多的细菌sRNA,开发了多个相关数据库。为了sRNA工作者系统了解与应用这些数据,本文拟对包含细菌sRNA的综合数据库和细菌sRNA专业数据库作一概述,并对sRNA数据库的未来发展进行展望。     

细菌非编码小RNA研究进展

细菌非编码小RNA(small non-coding RNA, sRNA)是一类长度在50~500个核苷酸, 不编码蛋白质的RNA。迄今, 在各种细菌中共发现超过150多种sRNA。它们通过碱基配对识别靶标mRNA, 在转录后水平调节基因的表达, 是细菌代谢、毒力和适应环境压力的重要调节因子。细菌sRNA的研究技术主要有基于生物信息学的计算机预测法和基于实验室的检测分析方法。这些方法所得到的sRNA都需要进行实验室确认, 然后再进一步通过各种实验手段研究其功能。     

非编码小RNA对细菌毒力及耐药性的调控作用研究进展

近年来的研究发现,细菌非编码小RNA (small non-coding RNA, sRNA)对其不同生理进程起到了重要的调控作用。随着大量sRNA被发现并鉴定,细菌sRNA的功能被逐步阐明,其可在转录后水平广泛调控细菌的生理代谢、毒力及耐药性等。本文综述了sRNA对细菌毒力和耐药性调控作用的研究进展,对揭示细菌转录后水平毒力及耐药性调控机制具有一定意义。     

细菌小RNA的调控及在代谢工程中的应用

细菌代谢工程需要优化基因的表达来平衡代谢物通量分布和减少有毒的中间体积累,从而提高产物生物合成。细菌小RNA(small RNA,sRNAs)与靶标mRNA通过碱基互补配对结合来抑制或激活其靶标基因的表达。sRNA在细菌的生理过程中都起到了至关重要的调控作用,因此被认为是细菌代谢工程中调节靶标基因表达的有力工具。近年来,越来越多的人工合成sRNA在细菌代谢工程中得到应用,分别就细菌sRNA的靶标识别和其对靶标的调控及代谢工程中的应用做了总结概括。     

Small RNA interactome of pathogenic E. coli revealed through crosslinking of RNase E

RNA sequencing studies have identified hundreds of non‐coding RNAs in bacteria, including regulatory small RNA (sRNA). However, our understanding of sRNA function has lagged behind their identification due to a lack of tools for the high‐throughput analysis of RNA–RNA interactions in bacteria. Here we demonstrate that in vivo sRNA–mRNA duplexes can be recovered using UV‐crosslinking, ligation and sequencing of hybrids (CLASH). Many sRNAs recruit the endoribonuclease, RNase E, to facilitate processing of mRNAs. We were able to recover base‐paired sRNA–mRNA duplexes in association with RNase E, allowing proximity‐dependent ligation and sequencing of cognate sRNA–mRNA pairs as chimeric reads. We verified that this approach captures bona fide sRNA–mRNA interactions. Clustering analyses identified novel sRNA seed regions and sets of potentially co‐regulated target mRNAs. We identified multiple mRNA targets for the pathotype‐specific sRNA Esr41, which was shown to regulate colicin sensitivity and iron transport in E. coli. Numerous sRNA interactions were also identified with non‐coding RNAs, including sRNAs and tRNAs, demonstrating the high complexity of the sRNA interactome.     

Identification of bacterial sRNA regulatory targets using ribosome profiling

Bacteria express large numbers of non-coding, regulatory RNAs known as ‘small RNAs’ (sRNAs). sRNAs typically regulate expression of multiple target messenger RNAs (mRNAs) through base-pairing interactions. sRNA:mRNA base-pairing often results in altered mRNA stability and/or altered translation initiation. Computational identification of sRNA targets is challenging due to the requirement for only short regions of base-pairing that can accommodate mismatches. Experimental approaches have been applied to identify sRNA targets on a genomic scale, but these focus only on those targets regulated at the level of mRNA stability. Here, we utilize ribosome profiling (Ribo-seq) to experimentally identify regulatory targets of the Escherichia coli sRNA RyhB. We not only validate a majority of known RyhB targets using the Ribo-seq approach, but also discover many novel ones. We further confirm regulation of a selection of known and novel targets using targeted reporter assays. By mutating nucleotides in the mRNA of a newly discovered target, we demonstrate direct regulation of this target by RyhB. Moreover, we show that Ribo-seq distinguishes between mRNAs regulated at the level of RNA stability and those regulated at the level of translation. Thus, Ribo-seq represents a powerful approach for genome-scale identification of sRNA targets.     

Major role for mRNA binding and restructuring in sRNA recruitment by Hfq

Bacterial small RNAs (sRNAs) modulate gene expression by base-pairing with target mRNAs. Many sRNAs require the Sm-like RNA binding protein Hfq as a cofactor. Well-characterized interactions between DsrA sRNA and the rpoS mRNA leader were used to understand how Hfq stimulates sRNA pairing with target mRNAs. DsrA annealing stimulates expression of rpoS by disrupting a secondary structure in the rpoS leader, which otherwise prevents translation. Both RNAs bind Hfq with similar affinity but interact with opposite faces of the Hfq hexamer. Using mutations that block interactions between two of the three components, we demonstrate that Hfq binding to a functionally critical (AAN)(4) motif in rpoS mRNA rescues DsrA binding to a hyperstable rpoS mutant. We also show that Hfq cannot stably bridge the RNAs. Persistent ternary complexes only form when the two RNAs are complementary. Thus, Hfq mainly acts by binding and restructuring the rpoS mRNA. However, Hfq binding to DsrA is needed for maximum annealing in vitro, indicating that transient interactions with both RNAs contribute to the regulatory mechanism.     

Quantitative effect of target translation on small RNA efficacy reveals a novel mode of interaction

Small regulatory RNAs (sRNAs) in bacteria regulate many important cellular activities under normal conditions and in response to stress. Many sRNAs bind to the mRNA targets at or near the 5′ untranslated region (UTR) resulting in translation inhibition and accelerated degradation. Often the sRNA-binding site is adjacent to or overlapping with the ribosomal binding site (RBS), suggesting a possible interplay between sRNA and ribosome binding. Here we combine quantitative experiments with mathematical modeling to reveal novel features of the interaction between small RNAs and the translation machinery at the 5′UTR of a target mRNA. By measuring the response of a library of reporter targets with varied RBSs, we find that increasing translation rate can lead to increased repression. Quantitative analysis of these data suggests a recruitment model, where bound ribosomes facilitate binding of the sRNA. We experimentally verified predictions of this model for the cell-to-cell variability of target expression. Our findings offer a framework for understanding sRNA silencing in the context of bacterial physiology.