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Cloning of a methanol-inducible moxF promoter and its analysis in moxB mutants of Methylobacterium extorquens AM1rif

Morris, Christina J. and Lidstrom, Mary E. (1992) Cloning of a methanol-inducible moxF promoter and its analysis in moxB mutants of Methylobacterium extorquens AM1rif. Journal of Bacteriology, 174 (13). pp. 4444-4449. ISSN 0021-9193. http://resolver.caltech.edu/CaltechAUTHORS:MORjbact92

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Abstract

In Methylobacterium extorquens AM1, genes encoding methanol dehydrogenase polypeptides are transcriptionally regulated in response to C, compounds, including methanol (M. E. Lidstrom and D. I. Stirling, Annu. Rev. Microbiol. 44:27-57, 1990). In order to study this regulation, a transcriptional fusion has been constructed between a beta-galactosidase reporter gene and a 1.55-kb XhoI-SalI fragment of M. extorquens AM1rif DNA encoding the N terminus of the methanol dehydrogenase large subunit (moxF) and 1,289 bp of upstream DNA. The fusion exhibited orientation-specific promoter activity in M. extorquens AM1rif but was expressed constitutively when the transcriptional fusion was located on the plasmid. However, correct regulation was restored when the construction was inserted in the M. extorquens AM1rif chromosome. This DNA fragment was shown to contain both the moxFJGI promoter and the sequences necessary in cis for its transcriptional regulation by methanol. Transcription from this promoter was studied in the M. extorquens AM1rif moxB mutant strains UV4rif and UV25rif, which have a pleiotropic phenotype with regard to the components of methanol oxidation. In these mutants, beta-galactosidase activity from the fusion was reduced to a level equal to that of the vector background when the fusion was present in both plasmid and chromosomal locations. Since both constitutive and methanol-inducible promoter activities were lost in the mutants, moxB appears to be required for transcription of the genes encoding the methanol dehydrogenase polypeptides.


Item Type:Article
Additional Information:Copyright © 1992, American Society for Microbiology. Received 24 January 1992. Accepted 21 April 1992. This work was supported by a grant from the Department of Energy (DE-FG03-87ER13753).
Subject Keywords:SP STRAIN AM1; DEHYDROGENASE STRUCTURAL GENE; NUCLEOTIDE-SEQUENCE; PARACOCCUS-DENITRIFICANS; OXIDATION; METHYLOTROPHS; EXPRESSION; PROTEINS
Record Number:CaltechAUTHORS:MORjbact92
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:MORjbact92
Alternative URL:http://www.pubmedcentral.gov/articlerender.fcgi?artid=206230
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:4266
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:10 Aug 2006
Last Modified:26 Dec 2012 08:58

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