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Giulia Oliva, Tobias Sahr, Monica Rolando, Maike Knoth, Carmen Buchrieser
: Legionella pneumophila is an environmental bacterium that parasitizes protozoa, but it may also infect humans, thereby causing a severe pneumonia called Legionnaires' disease. To cycle between the environment and a eukaryotic host, L. pneumophila is regulating the expression of virulence factors in a life cycle-dependent manner: replicating bacteria do not express virulence factors, whereas transmissive bacteria are highly motile and infective. Here we show that Hfq is an important regulator in this network...
January 10, 2017: MBio
Ashwin Lahiry, Samuel D Stimple, David W Wood, Richard A Lease
Multi-targeting small regulatory RNAs (sRNAs) represent a potentially useful tool for metabolic engineering applications. Natural multi-targeting sRNAs govern bacterial gene expression by binding to the translation initiation regions of protein-coding mRNAs through simple base pairing. We designed an Escherichia coli based genetic system to create and assay dual-acting retargeted-sRNA variants. The variants can be assayed for coordinate translational regulation of two alternate mRNA leaders fused to independent reporter genes...
January 9, 2017: ACS Synthetic Biology
Niko Popitsch, Ivana Bilusic, Philipp Rescheneder, Renée Schroeder, Meghan Lybecker
BACKGROUND: Transmission of Borrelia burgdorferi from its tick vector to a vertebrate host requires extensive reprogramming of gene expression. Small regulatory RNAs (sRNA) have emerged in the last decade as important regulators of bacterial gene expression. Despite the widespread observation of sRNA-mediated gene regulation, only one sRNA has been characterized in the Lyme disease spirochete B. burgdorferi. We employed an sRNA-specific deep-sequencing approach to identify the small RNA transcriptome of B...
January 5, 2017: BMC Genomics
Xiaoxia Ma, Ning Han, Chaogang Shao, Yijun Meng
Hints from animals point to the existence of two novel small RNA (sRNA) species surrounding the transcription start sites (TSSs) and the termini of the genes, respectively. In this study, we performed a comprehensive search for the two sRNA species named promoter-associated sRNAs (PASRs) and terminus-associated sRNAs (TASRs) in Arabidopsis. By using sRNA sequencing data from wild type plants and several mutants related to the sRNA biogenesis, Argonaute (AGO) 1- and AGO4-associated sRNA sequencing data, double-stranded RNA sequencing (dsRNA-seq) data, and DNA methylation profiling data, the biogenesis and action pathways of the PASRs and the TASRs were investigated...
2017: PloS One
Nan Li, Scott P Hennelly, Chris J Stubben, Sofiya Micheva-Viteva, Bin Hu, Yulin Shou, Momchilo Vuyisich, Chang-Shung Tung, Patrick S Chain, Karissa Y Sanbonmatsu, Elizabeth Hong-Geller
Non-coding small RNAs (sRNAs) are found in practically all bacterial genomes and play important roles in regulating gene expression to impact bacterial metabolism, growth, and virulence. We performed transcriptomics analysis to identify sRNAs that are differentially expressed in Yersinia pestis that invaded the human macrophage cell line THP-1, compared to pathogens that remained extracellular in the presence of host. Using ultra high-throughput sequencing, we identified 37 novel and 143 previously known sRNAs in Y...
2016: PloS One
Alberto Carbonell, José-Antonio Daròs
Artificial microRNAs (amiRNAs) and synthetic trans-acting small interfering RNAs (syn-tasiRNAs) are two classes of artificial small RNAs (sRNAs) engineered to silence endogenous transcripts as well as viral RNAs in plants. Here, we explore the possibility of using amiRNAs and syn-tasiRNAs to specifically interfere with infections by viroids, small (250-400 nt) non-coding circular RNAs with compact secondary structure infecting a wide range of plant species. The combined use of recent high-throughput methods for artificial sRNA construct generation and of the Potato spindle tuber viroid (PSTVd)/Nicotiana benthamiana pathosystem allowed for the simple and time-effective screening of multiple artificial sRNAs targeting sites distributed along PSTVd RNAs of (+) or (-) polarity...
December 27, 2016: Molecular Plant Pathology
Rachelle Bester, Johan T Burger, Hans J Maree
MicroRNAs (miRNAs) are a class of endogenous small non-coding RNAs (sRNA) that play an essential role in the regulation of target mRNAs expressed during plant development and in response to stress. MicroRNA expression profiling has helped to identify miRNAs that regulate a range of processes, including the plant's defence response to pathogens. In this study, differential miRNA expression in own-rooted Vitis vinifera cv. Cabernet Sauvignon plants infected with grapevine leafroll-associated virus 3 was investigated with microarrays and next-generation sequencing (NGS) of sRNA and mRNA...
December 26, 2016: Archives of Virology
Rongfang Guo, Yanping Deng, Zhongkai Huang, Xiaodong Chen, Xu XuHan, Zhongxiong Lai
MicroRNAs (miRNAs) are important for plant development including seed formation, dormancy, and germination, as well as seedling establishment. The Brassica vegetable seedling establishment stage influences the development of high quality seedlings, but also affects the nutrient content of sprouts. Chinese kale (Brassica alboglabra) seedlings at different growth stages were used to construct two small-RNA (sRNA) libraries. We comprehensively analyzed the miRNAs in 2- and 9-day-old seedlings. An average of 11,722,490 clean reads were generated after removing low-quality reads and adapter contaminants...
2016: Frontiers in Plant Science
Kook Han, Brian Tjaden, Stephen Lory
The first step in the post-transcriptional regulatory function of most bacterial small non-coding RNAs (sRNAs) is base pairing with partially complementary sequences of targeted transcripts. We present a simple method for identifying sRNA targets in vivo and defining processing sites of the regulated transcripts. The technique, referred to as global small non-coding RNA target identification by ligation and sequencing (GRIL-seq), is based on preferential ligation of sRNAs to the ends of base-paired targets in bacteria co-expressing T4 RNA ligase, followed by sequencing to identify the chimaeras...
December 22, 2016: Nature Microbiology
Raja Ragupathy, Sridhar Ravichandran, Md Safiur Rahman Mahdi, Douglas Huang, Elsa Reimer, Michael Domaratzki, Sylvie Cloutier
Understanding of plant adaptation to abiotic stresses has implications in plant breeding, especially in the context of climate change. MicroRNAs (miRNAs) and short interfering RNAs play a crucial role in gene regulation. Here, wheat plants were exposed to one of the following stresses: continuous light, heat or ultraviolet radiations over five consecutive days and leaf tissues from three biological replicates were harvested at 0, 1, 2, 3, 7 and 10 days after treatment (DAT). A total of 72 small RNA libraries were sequenced on the Illumina platform generating ~524 million reads corresponding to ~129 million distinct tags from which 232 conserved miRNAs were identified...
December 22, 2016: Scientific Reports
Feng Wang, Michael J Axtell
In plants, 24 nucleotide long heterochromatic siRNAs (het-siRNAs) transcriptionally regulate gene expression by RNA-directed DNA methylation (RdDM). The biogenesis of most het-siRNAs depends on the plant-specific RNA polymerase IV (Pol IV), and ARGONAUTE4 (AGO4) is a major het-siRNA effector protein. Through genome-wide analysis of sRNA-seq data sets, we found that AGO4 is required for the accumulation of a small subset of het-siRNAs. The accumulation of AGO4-dependent het-siRNAs also requires several factors known to participate in the effector portion of the RdDM pathway, including RNA POLYMERASE V (POL V), DOMAINS REARRANGED METHYLTRANSFERASE 2 (DRM2) and SAWADEE HOMEODOMAIN HOMOLOG 1 (SHH1)...
December 21, 2016: Plant Journal: for Cell and Molecular Biology
Chuang Shen, Ying-Ying Huang, Chun-Tao He, Qian Zhou, Jing-Xin Chen, Xiao Tan, Samavia Mubeen, Jian-Gang Yuan, Zhong-Yi Yang
In plants, microRNAs (miRNAs) play regulatory roles in response to various environmental stresses. In order to illustrate the regulation mechanisms of miRNAs involving the different Cd accumulation abilities between a low-shoot-Cd cultivar (QLQ) and a high-shoot-Cd cultivar (T308) of water spinach (Ipomoea aquatic Forsk.), six sRNA libraries at 3 different time points were constructed. Only 5 miRNAs were exclusively regulated in QLQ, among them, miRNA395 was up-regulated, which was supposed to enhance the Cd retention and detoxification in root...
December 12, 2016: Plant Physiology and Biochemistry: PPB
Trupti Joshi, Jiaojiao Wang, Hongxin Zhang, Shiyuan Chen, Shuai Zeng, Bowei Xu, Dong Xu
Soybean Knowledge Base (SoyKB) is a comprehensive all-inclusive web resource for bridging the gap between soybean translational genomics and molecular breeding. It provides information for six entities including genes/proteins, microRNAs (miRNAs)/small interfering RNAs (sRNA), metabolites, single nucleotide polymorphisms (SNPs), and plant introduction lines and traits. It has a user-friendly web interface publicly available at , which integrates and presents data in an intuitive manner to the soybean researchers, breeders, and consumers...
2017: Methods in Molecular Biology
Emiko Noro, Masaru Mori, Gakuto Makino, Yuki Takai, Sumiko Ohnuma, Asako Sato, Masaru Tomita, Kenji Nakahigashi, Akio Kanai
A new screening system for artificial small RNAs (sRNAs) that inhibit the growth of Escherichia coli was constructed. In this system, we used a plasmid library to express RNAs of ∼120 nucleotides, each with a random 30-nucleotide sequence that can recognize its target mRNA(s). After approximately 60,000 independent colonies were screened, several plasmids that inhibited bacterial growth were isolated. To understand the inhibitory mechanism, we focused on one sRNA, S-20, that exerted a strong inhibitory effect...
December 16, 2016: RNA Biology
Matthias Lechner, Mathias Schwarz, Madeleine Opitz, Erwin Frey
Post-transcriptional regulation of gene expression plays a crucial role in many bacterial pathways. In particular, the translation of mRNA can be regulated by trans-acting, small, non-coding RNAs (sRNAs) or mRNA-binding proteins, each of which has been successfully treated theoretically using two-component models. An important system that includes a combination of these modes of post-transcriptional regulation is the Colicin E2 system. DNA damage, by triggering the SOS response, leads to the heterogeneous expression of the Colicin E2 operon including the cea gene encoding the toxin colicin E2, and the cel gene that codes for the induction of cell lysis and release of colicin...
December 2016: PLoS Computational Biology
Paula J M van Kleeff, Marc Galland, Robert C Schuurink, Petra M Bleeker
The phloem-feeding whitefly Bemisia tabaci is a serious pest to a broad range of host plants, including many economically important crops such as tomato. These insects serve as a vector for various devastating plant viruses. It is known that whiteflies are capable of manipulating host-defense responses, potentially mediated by effector molecules in the whitefly saliva. We hypothesized that, beside putative effector proteins, small RNAs (sRNA) are delivered by B. tabaci into the phloem, where they may play a role in manipulating host plant defenses...
2016: Frontiers in Plant Science
Danielle L Michell, Ryan M Allen, Stuart R Landstreet, Shilin Zhao, Cynthia L Toth, Quanhu Sheng, Kasey C Vickers
The diversity of small non-coding RNAs (sRNA) is rapidly expanding and their roles in biological processes, including gene regulation, are emerging. Most interestingly, sRNAs are also found outside of cells and are stably present in all biological fluids. As such, extracellular sRNAs represent a novel class of disease biomarkers and are likely involved in cell signaling and intercellular communication networks. To assess their potential as biomarkers, sRNAs can be quantified in plasma, urine, and other fluids...
November 28, 2016: Journal of Visualized Experiments: JoVE
Yuzhe Sun, Zeta Mui, Xuan Liu, Aldrin Kay-Yuen Yim, Hao Qin, Fuk-Ling Wong, Ting-Fung Chan, Siu-Ming Yiu, Hon-Ming Lam, Boon Leong Lim
Small RNAs, including microRNAs (miRNAs) and phased small interfering RNAs (phasiRNAs; from PHAS loci), play key roles in plant development. Cultivated soybean, Glycine max, contributes a great deal to food production, but, compared to its wild kin, Glycine soja, it may lose some genetic information during domestication. In this work, we analyzed the sRNA profiles of different tissues in both cultivated (C08) and wild soybeans (W05) at three stages of development. A total of 443 known miRNAs and 15 novel miRNAs showed varying abundances between different samples, but the miRNA profiles were generally similar in both accessions...
December 6, 2016: International Journal of Molecular Sciences
Margarida Saramago, Alexandra Peregrina, Marta Robledo, Rute G Matos, Rolf Hilker, Javier Serrania, Anke Becker, Cecilia M Arraiano, José I Jiménez-Zurdo
Structural and biochemical features suggest that the almost ubiquitous bacterial YbeY protein may serve catalytic and/or Hfq-like protective functions central to small RNA (sRNA)-mediated regulation and RNA metabolism. We have biochemically and genetically characterized the YbeY ortholog of the legume symbiont Sinorhizobium meliloti (SmYbeY). Co-immunoprecipitation (CoIP) with a FLAG-tagged SmYbeY yielded a poor enrichment in RNA species, compared to Hfq CoIP-RNA uncovered previously by a similar experimental setup...
December 6, 2016: Nucleic Acids Research
Thao Nguyen Le Lam, Claire Morvan, Wenfeng Liu, Chantal Bohn, Yan Jaszczyszyn, Philippe Bouloc
Bacteria optimize their fitness in response to a changing environment by tight regulation of gene expression. Regulation can be controlled at both transcriptional and post-transcriptional levels via key players such as sigma factors, regulatory proteins and regulatory RNAs. The identification of phenotypes associated with gene deletions is the established method for finding gene functions but may require testing many conditions for each studied mutant. As regulatory RNAs often contribute to fine-tuning gene expression, phenotypes associated with their inactivation are often weak and difficult to detect...
December 2, 2016: Methods: a Companion to Methods in Enzymology
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