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1Department of Chemical Engineering, Box 351750, University of Washington, Seattle, WA 98195-1750, USA
2Department of Microbiology, Box 357242, University of Washington, Seattle, WA 98195-1750, USA
ABSTRACT
Five genes are thought to be required for transcription of methanol oxidation genes in Methylobacterium strains. These putative regulatory genes include mxcQE, which encode a putative sensor-regulator pair, and mxbDM and mxaB, whose functions are less well-understood. In this study, mxbDM in Methylobacterium extorquens AM1 were shown to be required for expression of a xyIE transcriptional fusion to the structural gene for the large subunit of methanol dehydrogenase (mxaF), confirming the role of these genes in transcriptional regulation of mxaF. The nucleotide sequence suggests that mxbD encodes a histidiine protein kinase with two transmembrane domains and that mxbM encodes a DNA-binding response regulator. A xyIE transcriptional fusion to the putative mxbD promoter showed low-level expression in wild-type cells grown on one-carbon (C1) compounds and no detectable expression in cells grown on succinate. Deletion analysis of this promoter construct showed that the region 229-129 bp upstream of the start of mxbD is required for expression. The expression of the mxbD-xylE fusion was examined in each of the five known regulatory mutant classes. xyIE expression was reduced to non-detectable levels in MxcQ and MxcE mutants, but was not affected in the other regulatory mutants or in non-regulatory mutants defective in methanol oxidation. These results suggest a regulatory hierarchy in which the sensor-regulator pair MxcQE control expression of the sensor-regulator pair MxbDM, and MxbDM in turn control expression of a number of genes involved in methanol oxidation.
Author for correspondence; Mary E. Lidstrom. Tel: +1 206 616 5282. Fax: +1 206 616 5721. e-mail: lidstrom@cheme.washington.edu
Present address: Department of Biochemistry, Hong Kong University of Science and Technology, Clear Water Bay Road, Kowloon, Hong Kong.
This article has been cited by other articles:
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  M. Zhang, K. A. FitzGerald, and M. E. Lidstrom Identification of an upstream regulatory sequence that mediates the transcription of mox genes in Methylobacterium extorquens AM1 Microbiology, November 1, 2005; 151(11): 3723 - 3728. [Abstract] [Full Text] [PDF]
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M. Zhang and M. E. Lidstrom Promoters and transcripts for genes involved in methanol oxidation in Methylobacterium extorquens AM1 Microbiology, April 1, 2003; 149(4): 1033 - 1040. [Abstract] [Full Text] [PDF]
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