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Klein, Thomas ; Funke, Franziska ; Rossbach, Oliver ; Lehmann, Gerhard ; Vockenhuber, Michael ; Medenbach, Jan ; Suess, Beatrix ; Meister, Gunter ; Babinger, Patrick

Investigating the Prevalence of RNA-Binding Metabolic Enzymes in E. coli

Klein, Thomas, Funke, Franziska , Rossbach, Oliver , Lehmann, Gerhard, Vockenhuber, Michael, Medenbach, Jan , Suess, Beatrix, Meister, Gunter und Babinger, Patrick (2023) Investigating the Prevalence of RNA-Binding Metabolic Enzymes in E. coli. International Journal of Molecular Sciences 24 (14), S. 11536.

Veröffentlichungsdatum dieses Volltextes: 20 Jul 2023 07:46
Artikel
DOI zum Zitieren dieses Dokuments: 10.5283/epub.54500


Zusammenfassung

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 ...

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.



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  • [img] [img] Klein, Thomas, Funke, Franziska , Rossbach, Oliver , Lehmann, Gerhard, Vockenhuber, Michael, Medenbach, Jan , Suess, Beatrix, Meister, Gunter und Babinger, Patrick (2023) Investigating the Prevalence of RNA-Binding Metabolic Enzymes in E. coli. International Journal of Molecular Sciences 24 (14), S. 11536. [Gegenwärtig angezeigt]

Details

DokumentenartArtikel
Titel eines Journals oder einer ZeitschriftInternational Journal of Molecular Sciences
Verlag:MDPI
Ort der Veröffentlichung:BASEL
Band:24
Nummer des Zeitschriftenheftes oder des Kapitels:14
Seitenbereich:S. 11536
Datum16 Juli 2023
InstitutionenBiologie und Vorklinische Medizin > Institut für Biophysik und physikalische Biochemie
Biologie und Vorklinische Medizin > Institut für Biochemie, Genetik und Mikrobiologie
Identifikationsnummer
WertTyp
10.3390/ijms241411536DOI
Stichwörter / KeywordsAU-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
Dewey-Dezimal-Klassifikation500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie
500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie
StatusVeröffentlicht
BegutachtetJa, diese Version wurde begutachtet
An der Universität Regensburg entstandenJa
URN der UB Regensburgurn:nbn:de:bvb:355-epub-545003
Dokumenten-ID54500

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