MreA Functions in the Global Regulation of Methanogenic Pathways in <named-content content-type="genus-species">Methanosarcina acetivorans</named-content>

ABSTRACT Results are presented supporting a regulatory role for the product of the MA3302 gene locus (designated MreA) previously annotated as a hypothetical protein in the methanogenic species Methanosarcina acetivorans of the domain Archaea. Sequence analysis of MreA revealed identity to the TrmB...

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Autores principales:	Matthew J. Reichlen, Venkata R. Vepachedu, Katsuhiko S. Murakami, James G. Ferry
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2012
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/680fdbee9f1d489cadeb888526296b21
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spelling	oai:doaj.org-article:680fdbee9f1d489cadeb888526296b212021-11-15T15:39:09ZMreA Functions in the Global Regulation of Methanogenic Pathways in <named-content content-type="genus-species">Methanosarcina acetivorans</named-content>10.1128/mBio.00189-122150-7511https://doaj.org/article/680fdbee9f1d489cadeb888526296b212012-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00189-12https://doaj.org/toc/2150-7511ABSTRACT Results are presented supporting a regulatory role for the product of the MA3302 gene locus (designated MreA) previously annotated as a hypothetical protein in the methanogenic species Methanosarcina acetivorans of the domain Archaea. Sequence analysis of MreA revealed identity to the TrmB family of transcription factors, albeit the sequence is lacking the sensor domain analogous to TrmBL2, abundant in nonmethanogenic species of the domain Archaea. Transcription of mreA was highly upregulated during growth on acetate versus methylotrophic substrates, and an mreA deletion (ΔmreA) strain was impaired for growth with acetate in contrast to normal growth with methylotrophic substrates. Transcriptional profiling of acetate-grown cells identified 280 genes with altered expression in the ΔmreA strain versus the wild-type strain. Expression of genes unique to the acetate pathway decreased whereas expression of genes unique to methylotrophic metabolism increased in the ΔmreA strain relative to the wild type, results indicative of a dual role for MreA in either the direct or indirect activation of acetate-specific genes and repression of methylotrophic-specific genes. Gel shift experiments revealed specific binding of MreA to promoter regions of regulated genes. Homologs of MreA were identified in M. acetivorans and other Methanosarcina species for which expression patterns indicate roles in regulating methylotrophic pathways. IMPORTANCE Species in the domain Archaea utilize basal transcription machinery resembling that of the domain Eukarya, raising questions addressing the role of numerous putative transcription factors identified in sequenced archaeal genomes. Species in the genus Methanosarcina are ideally suited for investigating principles of archaeal transcription through analysis of the capacity to utilize a diversity of substrates for growth and methanogenesis. Methanosarcina species switch pathways in response to the most energetically favorable substrate, metabolizing methylotrophic substrates in preference to acetate marked by substantial regulation of gene expression. Although conversion of the methyl group of acetate accounts for most of the methane produced in Earth’s biosphere, no proteins involved in the regulation of genes in the acetate pathway have been reported. The results presented here establish that MreA participates in the global regulation of diverse methanogenic pathways in the genus Methanosarcina. Finally, the results contribute to a broader understanding of transcriptional regulation in the domain Archaea.Matthew J. ReichlenVenkata R. VepacheduKatsuhiko S. MurakamiJames G. FerryAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 3, Iss 4 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Matthew J. Reichlen Venkata R. Vepachedu Katsuhiko S. Murakami James G. Ferry MreA Functions in the Global Regulation of Methanogenic Pathways in <named-content content-type="genus-species">Methanosarcina acetivorans</named-content>
description	ABSTRACT Results are presented supporting a regulatory role for the product of the MA3302 gene locus (designated MreA) previously annotated as a hypothetical protein in the methanogenic species Methanosarcina acetivorans of the domain Archaea. Sequence analysis of MreA revealed identity to the TrmB family of transcription factors, albeit the sequence is lacking the sensor domain analogous to TrmBL2, abundant in nonmethanogenic species of the domain Archaea. Transcription of mreA was highly upregulated during growth on acetate versus methylotrophic substrates, and an mreA deletion (ΔmreA) strain was impaired for growth with acetate in contrast to normal growth with methylotrophic substrates. Transcriptional profiling of acetate-grown cells identified 280 genes with altered expression in the ΔmreA strain versus the wild-type strain. Expression of genes unique to the acetate pathway decreased whereas expression of genes unique to methylotrophic metabolism increased in the ΔmreA strain relative to the wild type, results indicative of a dual role for MreA in either the direct or indirect activation of acetate-specific genes and repression of methylotrophic-specific genes. Gel shift experiments revealed specific binding of MreA to promoter regions of regulated genes. Homologs of MreA were identified in M. acetivorans and other Methanosarcina species for which expression patterns indicate roles in regulating methylotrophic pathways. IMPORTANCE Species in the domain Archaea utilize basal transcription machinery resembling that of the domain Eukarya, raising questions addressing the role of numerous putative transcription factors identified in sequenced archaeal genomes. Species in the genus Methanosarcina are ideally suited for investigating principles of archaeal transcription through analysis of the capacity to utilize a diversity of substrates for growth and methanogenesis. Methanosarcina species switch pathways in response to the most energetically favorable substrate, metabolizing methylotrophic substrates in preference to acetate marked by substantial regulation of gene expression. Although conversion of the methyl group of acetate accounts for most of the methane produced in Earth’s biosphere, no proteins involved in the regulation of genes in the acetate pathway have been reported. The results presented here establish that MreA participates in the global regulation of diverse methanogenic pathways in the genus Methanosarcina. Finally, the results contribute to a broader understanding of transcriptional regulation in the domain Archaea.
format	article
author	Matthew J. Reichlen Venkata R. Vepachedu Katsuhiko S. Murakami James G. Ferry
author_facet	Matthew J. Reichlen Venkata R. Vepachedu Katsuhiko S. Murakami James G. Ferry
author_sort	Matthew J. Reichlen
title	MreA Functions in the Global Regulation of Methanogenic Pathways in <named-content content-type="genus-species">Methanosarcina acetivorans</named-content>
title_short	MreA Functions in the Global Regulation of Methanogenic Pathways in <named-content content-type="genus-species">Methanosarcina acetivorans</named-content>
title_full	MreA Functions in the Global Regulation of Methanogenic Pathways in <named-content content-type="genus-species">Methanosarcina acetivorans</named-content>
title_fullStr	MreA Functions in the Global Regulation of Methanogenic Pathways in <named-content content-type="genus-species">Methanosarcina acetivorans</named-content>
title_full_unstemmed	MreA Functions in the Global Regulation of Methanogenic Pathways in <named-content content-type="genus-species">Methanosarcina acetivorans</named-content>
title_sort	mrea functions in the global regulation of methanogenic pathways in <named-content content-type="genus-species">methanosarcina acetivorans</named-content>
publisher	American Society for Microbiology
publishDate	2012
url	https://doaj.org/article/680fdbee9f1d489cadeb888526296b21
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MreA Functions in the Global Regulation of Methanogenic Pathways in <named-content content-type="genus-species">Methanosarcina acetivorans</named-content>

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