MnoSR removal in Mycobacterium smegmatis triggers broad transcriptional response to 1,3-propanediol and glucose as sole carbon sources.

Autor:	Płocińska R; Institute of Medical Biology of the Polish Academy of Sciences, Łódź, Poland., Struś K; Institute of Medical Biology of the Polish Academy of Sciences, Łódź, Poland., Korycka-Machała M; Institute of Medical Biology of the Polish Academy of Sciences, Łódź, Poland., Płociński P; Department of Immunology and Infectious Biology, Faculty of Biology and Environmental Protection, University of Łódz, Łódź, Poland., Kuzioła M; Institute of Medical Biology of the Polish Academy of Sciences, Łódź, Poland.; BioMedChem Doctoral School of the UL and Łódź Institutes of the Polish Academy of Sciences, Łódź, Poland., Żaczek A; Department of Microbiology, College of Medical Sciences, University of Rzeszów, Rzeszów, Poland., Słomka M; Biobank Lab, Department of Molecular Biophysics, Faculty of Biology and Environmental Protection, University of Łódź, Łódź, Poland., Dziadek J; Institute of Medical Biology of the Polish Academy of Sciences, Łódź, Poland.
Jazyk:	angličtina
Zdroj:	Frontiers in cellular and infection microbiology [Front Cell Infect Microbiol] 2024 Jul 24; Vol. 14, pp. 1427829. Date of Electronic Publication: 2024 Jul 24 (Print Publication: 2024).
DOI:	10.3389/fcimb.2024.1427829
Abstrakt:	Introduction: The two-component signal transduction systems play an essential role in the adaptation of bacteria to changing environmental conditions. One of them is the MnoSR system involved in the regulation of methylotrophic metabolism in M. smegmatis. Methods: Mycobacterium smegmatis mutant strains ΔmnoS, ΔmnoR and ΔmnoS/R lacking functional mnoS, mnoR and both genes were generated using a homologous recombination approach. MnoR recombinant protein was purified by affinity column chromatography. The present study employs molecular biology techniques: cloning strategies, global RNA sequencing, qRT-PCR, EMSA, Microscale thermophoresis, and bioinformatics analysis. Results and Discussion: The ∆mnoS, ∆mnoR, and ∆mnoS/R mutant strains were generated and cultured in the presence of defined carbon sources. Growth curve analysis confirmed that inactivation of the MnoSR impairs the ability of M. smegmatis cells to use alcohols such as 1,3-propanediol and ethanol but improves the bacterial growth on ethylene glycol, xylitol, and glycerol. The total RNA sequencing method was employed to understand the importance of MnoSR in the global responses of mycobacteria to limited carbon access and in carbon-rich conditions. The loss of MnoSR significantly affected carbon utilization in the case of mycobacteria cultured on glucose or 1,3-propanediol as sole carbon sources as it influenced the expression of multiple metabolic pathways. The numerous transcriptional changes could not be linked to the presence of evident MnoR DNA-binding sites within the promotor regions for the genes outside of the mno operon. This was confirmed by EMSA and microscale thermophoresis with mutated MnoR binding consensus region. Our comprehensive analysis highlights the system's vital role in metabolic adaptability, providing insights into its potential impact on the environmental survival of mycobacteria. Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2024 Płocińska, Struś, Korycka-Machała, Płociński, Kuzioła, Żaczek, Słomka and Dziadek.)
Databáze:	MEDLINE
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