Klein, Thomas ; Funke, Franziska ; Rossbach, Oliver ; Lehmann, Gerhard ; Vockenhuber, Michael ; Medenbach, Jan ; Suess, Beatrix ; Meister, Gunter ; Babinger, Patrick (2023)
Investigating the Prevalence of RNA-Binding Metabolic Enzymes in E. coli.
In: International Journal of Molecular Sciences, 2023, 24 (14)
doi: 10.26083/tuprints-00024407
Artikel, Zweitveröffentlichung, Verlagsversion
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Kurzbeschreibung (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.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2023 |
| Autor(en): | Klein, Thomas ; Funke, Franziska ; Rossbach, Oliver ; Lehmann, Gerhard ; Vockenhuber, Michael ; Medenbach, Jan ; Suess, Beatrix ; Meister, Gunter ; Babinger, Patrick |
| Art des Eintrags: | Zweitveröffentlichung |
| Titel: | Investigating the Prevalence of RNA-Binding Metabolic Enzymes in E. coli |
| Sprache: | Englisch |
| Publikationsjahr: | 2023 |
| Ort: | Darmstadt |
| Publikationsdatum der Erstveröffentlichung: | 2023 |
| Verlag: | MDPI |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | International Journal of Molecular Sciences |
| Jahrgang/Volume einer Zeitschrift: | 24 |
| (Heft-)Nummer: | 14 |
| Kollation: | 23 Seiten |
| DOI: | 10.26083/tuprints-00024407 |
| URL / URN: | https://tuprints.ulb.tu-darmstadt.de/24407 |
| Zugehörige Links: | |
| Herkunft: | Zweitveröffentlichung DeepGreen |
| Kurzbeschreibung (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. |
| Freie Schlagworte: | metabolic enzymes, REM hypothesis, RNA-binding protein, prokaryotes, SELEX, unconventional RNA binding, MS2 phage, quinone oxidoreductase |
| Status: | Verlagsversion |
| URN: | urn:nbn:de:tuda-tuprints-244075 |
| Zusätzliche Informationen: | This article belongs to the Special Issue Small Prokaryotic Proteins Interacting with Nucleic Acids 2.0 |
| Sachgruppe der Dewey Dezimalklassifikatin (DDC): | 500 Naturwissenschaften und Mathematik > 540 Chemie 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie |
| Fachbereich(e)/-gebiet(e): | Interdisziplinäre Forschungsprojekte Interdisziplinäre Forschungsprojekte > Centre for Synthetic Biology |
| Hinterlegungsdatum: | 11 Aug 2023 12:09 |
| Letzte Änderung: | 14 Aug 2023 08:52 |
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| Suche nach Titel in: | TUfind oder in Google |
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