The study on Fnr-type transcription regulators functions in bacterium Paracoccus denitrificans using proteomics, transcriptomics and bioinformatics tools

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Authors

BOUCHAL Pavel STRUHÁROVÁ Iva BUDINSKÁ Eva ŠEDO Ondrej VYHLÍDALOVÁ Tereza ZDRÁHAL Zbyněk VAN SPANNING Rob KUČERA Igor

Year of publication 2009
Type Conference abstract
MU Faculty or unit

Faculty of Science

Citation
Description Paracoccus denitrificans is a facultatively autotrophic soil bacterium which responds to decreasing level of oxygen in the environment by a switch from aerobic to anaerobic (denitrifying) growth mode. In our work, we analyzed roles of transcription regulators FnrP, NNR and NarR (with described key roles as sensors for oxygen, nitrite and nitric oxide) using proteomic, transcriptomic and bioinformatics tools to consider their roles in the adaptation processes at global level. Protein compositions of four P. denitrificans strains (wild type, FnrP-, NNR- and NarR- mutants grown aerobically, semiaerobically and semiaerobically with nitrate) were analyzed using set of 36 large format 2-D PAGE gels, their statistical evaluation and MS/MS protein identification of more than 500 proteins. Expression differences of key up-/down-regulated gene products were validated using qRT-PCR. Comparative proteomic analysis revealed differences between strains in the key proteins involved in denitrification (nitrate reductase, nitrite reductase, nitrous oxide reductase) and OmpW and UspA proteins. These findings are in agreement with previously published data and confirmed the positions of proposed fnr binding sites in their promoters. Detection of additional group of up-/down- regulated proteins (without fnr-binding sites in their promoters) indicates involvement of additional regulatory mechanisms into the adaptation processes studied. Evidences for potential roles of transcription regulators of other families than Fnr are discussed.
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