Regulation of putative methyl-sulphide methyltransferases in Methanosarcina acetivorans C2A

The regulation of the Methanosarcina acetivorans mtsD, mtsF and mtsH genes, which encode putative corrinoid/methyltransferase isozymes involved in methylsulphide metabolism, was examined by a variety of methods, suggesting that their expression is regulated at both the transcriptional and post-trans...

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Published in:Molecular microbiology 2009-10, Vol.74 (1), p.227-238
Main Authors: Bose, Arpita, Kulkarni, Gargi, Metcalf, William W
Format: Article
Language:English
Subjects:
Archaeal Proteins - genetics
Archaeal Proteins - metabolism
Bacteriology
Base Sequence
Biological and medical sciences
Fundamental and applied biological sciences. Psychology
Gene expression
Gene Expression Regulation, Archaeal
Genes, Archaeal
Methanol
Methanosarcina - enzymology
Methanosarcina - genetics
Methyltransferases - genetics
Methyltransferases - metabolism
Microbiology
Miscellaneous
Molecular Sequence Data
Mutation
Promoter Regions, Genetic
RNA, Archaeal - genetics
RNA, Messenger - genetics
Substrate Specificity
Transcription Initiation Site
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Summary:The regulation of the Methanosarcina acetivorans mtsD, mtsF and mtsH genes, which encode putative corrinoid/methyltransferase isozymes involved in methylsulphide metabolism, was examined by a variety of methods, suggesting that their expression is regulated at both the transcriptional and post-transcriptional levels. Transcripts of all three genes, measured by quantitative reverse transcription PCR, were shown to be most abundant during growth on methanol with dimethylsulphide (DMS). Transcript levels were also high in media with CO or methylamines, but much lower with methanol. In contrast, translational fusions to mtsD showed high expression levels on CO or methanol with DMS, while the mtsF translational fusion showed highest reporter gene activity on methylamines with much lower expression on CO or methanol with DMS. The activity of mtsD and mtsF fusions was very low when the strains were grown in methanol or acetate. Expression of the mtsH fusion was not detected on any substrate, despite the presence of an mRNA transcript. The transcription start sites of all three genes were determined by 5'-RACE revealing large leader sequences for each transcript. Characterization of deletion mutants lacking putative regulatory genes suggests that MA0862 (msrF), MA4383 (msrC) and MA4560 (msrG) act as transcriptional activators of mtsD, mtsF and mtsH respectively.
ISSN:0950-382X
1365-2958
DOI:10.1111/j.1365-2958.2009.06864.x