@article{10902/36120,
year = {2021},
url = {https://hdl.handle.net/10902/36120},
abstract = {Function of bacterial small non-coding RNAs (sRNAs) and overall RNA metabolism is largely shaped by a vast diversity of RNA-protein interactions. However, in non-model bacteria with defined non-coding transcriptomes the sRNA interactome remains almost unexplored. We used affinity chromatography to capture proteins associated in vivo with MS2-tagged trans-sRNAs that regulate nutrient uptake (AbcR2 and NfeR1) and cell cycle (EcpR1) mRNAs by antisense-based translational inhibition in the nitrogen-fixing α-rhizobia Sinorhizobium meliloti. The three proteomes were rather distinct, with that of EcpR1 particularly enriched in cell cycle-related enzymes, whilst sharing several transcription/translation-related proteins recurrently identified associated with sRNAs. Strikingly, MetK, the synthetase of the major methyl donor S-adenosylmethionine, was reliably recovered as a binding partner of the three sRNAs, which reciprocally co-immunoprecipitated with a FLAG-tagged MetK variant. Induced (over)expression of the trans-sRNAs and MetK depletion did not influence canonical riboregulatory traits, `for example, protein titration or sRNA stability, respectively. An in vitro filter assay confirmed binding of AbcR2, NfeR1 and EcpR1 to MetK and further revealed interaction of the protein with other non-coding and coding transcripts but not with the 5S rRNA. These findings uncover a broad specificity for RNA binding as an unprecedented feature of this housekeeping prokaryotic enzyme.},
organization = {Funding: This work was supported by the Spanish Ministerio de Ciencia, Innovación y Universidades under ERDF-cofinanced grants [BFU2013-48282-C2-2-P and BFU2017-82645-P to J.I.J.-Z.], a contract of the program ‘Formación Post-doctoral’ (Juan de la Cierva) [FPDI-2013-16255 to M.R.], and an FPU fellowship [FPU16/01275 to N.I.G.-T.]. Acknowledgments
We thank Prof. Henning Urlaub (Institute for Clinical Chemistry, University Medical Center Göttingen) for kindly providing the plasmid for MS2-MBP expression. The proteomic analyses were performed in the Proteomic Unit of IPBLN-CSIC that belongs to ProteoRed, PRB2-ISCIII,
supported by grant PT13/0001.},
publisher = {Taylor & Francis},
publisher = {RNA Biology, 2024, 18(8), 1111-1123},
title = {Synthetase of the methyl donor S-adenosylmethionine from nitrogen-fixing a-rhizobia can bind functionally diverse RNA species},
author = {Robledo Garrido, Marta and García Tomsig, Natalia I. and Matia González, Ana M. and García Rodríguez, Fernando M. and Jiménez Zurdo, José I.},
}