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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 and Babinger, Patrick (2023) Investigating the Prevalence of RNA-Binding Metabolic Enzymes in E. coli. International Journal of Molecular Sciences 24 (14), p. 11536.

Date of publication of this fulltext: 20 Jul 2023 07:46
Article
DOI to cite this document: 10.5283/epub.54500


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 ...

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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Details

Item typeArticle
Journal or Publication TitleInternational Journal of Molecular Sciences
Publisher:MDPI
Open Access Type:Gold (with APC)
Place of Publication:BASEL
Volume:24
Number of Issue or Book Chapter:14
Page Range:p. 11536
Date16 July 2023
InstitutionsBiology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie
Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie
Identification Number
ValueType
10.3390/ijms241411536DOI
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 Decimal Classification500 Science > 570 Life sciences
500 Science > 570 Life sciences
StatusPublished
RefereedYes, this version has been refereed
Created at the University of RegensburgYes
URN of the UB Regensburgurn:nbn:de:bvb:355-epub-545003
Item ID54500

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