Klein, Thomas and Funke, Franziska and Rossbach, Oliver and Lehmann, Gerhard and Vockenhuber, Michael and Medenbach, Jan and Suess, Beatrix and Meister, Gunter and Babinger, Patrick (2023) Investigating the Prevalence of RNA-Binding Metabolic Enzymes in <i>E. coli</i>. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 24 (14): 11536. ISSN 1661-6596, 1422-0067
Full text not available from this repository. (Request a copy)Abstract
An open research field in cellular regulation is the assumed crosstalk between RNAs, metabolic enzymes, and metabolites, also known as the REM hypothesis. High-throughput assays have produced extensive interactome data with metabolic enzymes frequently found as hits, but only a few examples have been biochemically validated, with deficits especially in prokaryotes. Therefore, we rationally selected nineteen Escherichia coli enzymes from such datasets and examined their ability to bind RNAs using two complementary methods, iCLIP and SELEX. Found interactions were validated by EMSA and other methods. For most of the candidates, we observed no RNA binding (12/19) or a rather unspecific binding (5/19). Two of the candidates, namely glutamate-5-kinase (ProB) and quinone oxidoreductase (QorA), displayed specific and previously unknown binding to distinct RNAs. We concentrated on the interaction of QorA to the mRNA of yffO, a grounded prophage gene, which could be validated by EMSA and MST. Because the physiological function of both partners is not known, the biological relevance of this interaction remains elusive. Furthermore, we found novel RNA targets for the MS2 phage coat protein that served us as control. Our results indicate that RNA binding of metabolic enzymes in procaryotes is less frequent than suggested by the results of high-throughput studies, but does occur.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | AU-RICH RNA; ESCHERICHIA-COLI; MESSENGER-RNA; GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE; IN-VITRO; CRYSTAL-STRUCTURE; ZETA-CRYSTALLIN; POSTTRANSCRIPTIONAL REGULATION; ADENYLATE KINASE; CROSS-LINKING; metabolic enzymes; REM hypothesis; RNA-binding protein; prokaryotes; SELEX; unconventional RNA binding; MS2 phage; quinone oxidoreductase |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 08 Mar 2024 14:06 |
| Last Modified: | 08 Mar 2024 14:06 |
| URI: | https://pred.uni-regensburg.de/id/eprint/59191 |
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