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BMC Genomics

, 18:343

Prokaryote microbial genomics

Abstract

BackgroundBacterial small regulatory RNAs sRNAs play important roles in sensing environment changes through sRNA-target mRNA interactions. However, the current strategy for detecting sRNA-mRNA interactions usually combines bioinformatics prediction and experimental verification, which is hampered by low prediction accuracy and low-throughput. Additionally, among the 4736 sequenced bacterial genomes, only about 2164 sRNAs from 319 strains have been described. Furthermore, target mRNAs of only 157 sRNAs have been uncovered. Obviously, highly efficient methods were required to detect sRNA-mRNA interactions in the sequenced genomes. This study aimed to apply a modified CLASH cross-linking, ligation and sequencing hybrids method to detect RNA-RNA interactions in E. coli, a model bacterial organism.

ResultsStatistically significant interactions were detected in 29 transcript pairs. To the best of our knowledge, 24 pairs were reported for the first time and were novel RNA interactions, including tRNA-tRNA, tRNA-ncRNA non-coding RNA, tRNA-rRNA, rRNA-mRNA, rRNA-ncRNA, rRNA-rRNA, rRNA-IGT intergenic transcript, and tRNA-IGT interactions.

ConclusionsDiscovery of novel RNA-RNA interactions in the present study demonstrates that RNA-RNA interactions might be far more complicated than ever expected. New methods may be required to help discover more novel RNA-RNA interactions. The present work describes a high-throughput protocol not only for discovering new RNA interactions, but also directly obtaining base-pairing sequences, which should be useful in assessing RNA structure and interactions.

KeywordsRNA-RNA interaction High-throughput sequencing Bacteria AbbreviationsAMT4’-aminomethyl-trioxsalen

CLASHCross-linking, ligation and sequencing hybrids

HITSHigh-throughput sequencing

HITS-CLIPHigh-throughput sequencing of RNAs isolated by crosslinking immunoprecipitation

IGTIntergenic transcript

iPAR-CLIPIn vivo PAR-CLIP

ncRNANon-coding RNA

PAR-CLIPPhotoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation

PNKPolynucleotide kinase

RRIRNA-RNA interaction

sRNASmall regulatory RNA

tmRNATransfer-messenger RNA

Electronic supplementary materialThe online version of this article doi:10.1186-s12864-017-3725-3 contains supplementary material, which is available to authorized users.





Autor: Tao Liu - Kaiyu Zhang - Song Xu - Zheng Wang - Hanjiang Fu - Baolei Tian - Xiaofei Zheng - Wuju Li

Fuente: https://link.springer.com/

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