期刊
METHODS
卷 63, 期 2, 页码 135-143出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2013.06.033
关键词
Endoribonuclease; Antisense RNA regulation; Regulatory RNAs; RNA-RNA interactions; RNA-protein interaction
资金
- Centre National de la Recherche Scientifique (CNRS)
- Agence Nationale de la Recherche [ANR10-Pathogenomics-ARMSA]
- Laboratoires d'excellence (LABEX) NetRNA grant in the frame of Programme d'Investissements d'Avenir, the Ministry of Science and Technology (Israel) [ANR-10-LABX-36]
- Ministry of Foreign and European Affairs
- Ministry of Higher Education and Research (France) in the frame of a network program Complexity in Biology
Ribonucleases play key roles in gene regulation and in the expression of virulence factors in Staphylococcus aureus. Among these enzymes, the double-strand specific endoribonuclease III (RNase III) is a key mediator of mRNA processing and degradation. Recently, we have defined, direct target sites for RNase III processing on a genorne-wide scale in S. aureus. Our approach is based on deep sequencing of cDNA libraries obtained from RNAs isolated by in vivo co-immunoprecipitation with wild-type RNase III and two cleavage-defective mutants. The use of such catalytically inactivated enzymes, which still retain their RNA binding capacity; allows the identification of novel RNA substrates of RNase III. In this report, we will summarize the diversity of RNase III functions, discuss the advantages and the limitations of the approach, and how this strategy identifies novel mRNA targets of small non-coding RNAs in S. aureus. (C) 2013 Elsevier Inc. All rights reserved.
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