Identification of an upstream regulatory sequence that mediates the transcription of mox genes in Methylobacterium extorquens AM1
- 1 November 2005
- journal article
- Published by Microbiology Society in Microbiology
- Vol. 151 (11) , 3723-3728
- https://doi.org/10.1099/mic.0.28243-0
Abstract
A multiple A-tract sequence has been identified in the promoter regions for the mxaF, pqqA, mxaW, mxbD and mxcQ genes involved in methanol oxidation in Methylobacterium extorquens AM1, a facultative methylotroph. Site-directed mutagenesis was exploited to delete or change this conserved sequence. Promoter-xylE transcriptional fusions were used to assess promoter activity in these mutants. A fiftyfold drop in the XylE activity was observed for the mxaF and pqqA promoters without this sequence, and a five- to sixfold drop in the XylE activity was observed for the mxbD and mxcQ promoters without this sequence. Mutants were generated in the chromosomal copies in which this sequence was either deleted or altered, and these mutants were unable to grow on methanol. When one of these sequences was added to Plac of Escherichia coli, which is a weak constitutive promoter in M. extorquens AM1, the activity increased two- to threefold. These results suggest that this sequence is essential for normal expression of these genes in M. extorquens AM1, and may serve as a general enhancer element for genetic constructs in this bacterium.Keywords
This publication has 32 references indexed in Scilit:
- Methylotrophy in Methylobacterium extorquens AM1 from a Genomic Point of ViewJournal of Bacteriology, 2003
- Promoters and transcripts for genes involved in methanol oxidation in Methylobacterium extorquens AM1Microbiology, 2003
- Novel Methylotrophy Genes of Methylobacterium extorquens AM1 Identified by using Transposon Mutagenesis Including a Putative Dihydromethanopterin ReductaseJournal of Bacteriology, 2003
- Sequence and characterization ofmxaB, a response regulator involved in regulation of methanol oxidation, and ofmxaW, a methanol-regulated gene inMethylobacterium extorquensAM1FEMS Microbiology Letters, 1998
- Molecular analysis of mxbD and mxbM, a putative sensor-regulator pair required for oxidation of methanol in Methylobacterium extorquens AM1Microbiology, 1997
- Sequence analysis of pqq genes required for biosynthesis of pyrroloquinoline quinone in Methylobacterium extorquens AM1 and the purification of a biosynthetic intermediateMicrobiology, 1997
- Methanol oxidation mutants in Methylobacterium extorquens AM1: identification of new genetic complementation groupsMicrobiology, 1995
- Nucleotide sequence of the Methylobacterium extorquens AM1 moxF and moxJ genes involved in methanol oxidationGene, 1990
- Chromogenic identification of genetic regulatory signals in Bacillus subtilis based on expression of a cloned Pseudomonas gene.Proceedings of the National Academy of Sciences, 1983
- Preparation of transforming deoxyribonucleic acid by phenol treatmentBiochimica et Biophysica Acta (BBA) - Specialized Section on Nucleic Acids and Related Subjects, 1963