Abstract
The organization of genes involved in utilization of methylamine (mau genes) was studied in Methylophilus methylotrophus W3A1. The strain used was a nonmucoid variant termed NS (nonslimy). The original mucoid strain was shown to be identical to the NS strains on the basis of chromosomal digest and hybridization patterns. An 8-kb PstI fragment of the chromosome from M. methylotrophus W3A1-NS encoding the mau genes was cloned and a 6,533-bp region was sequenced. Eight open reading frames were found inside the sequenced area. On the basis of a high level of sequence identity with the Mau polypeptides from Methylobacterium extorquens AM1, the eight open reading frames were identified as mauFBEDAGLM. The mau gene cluster from M. methylotrophus W3A1 is missing two genes, mauC (amicyanin) and mauJ (whose function is unknown), which have been found between mauA and mauG in all studied mau gene clusters. Mau polypeptides sequenced so far from five different bacteria show considerable identity. A mauA mutant of M. methylotrophus W3A1-NS that was constructed lost the ability to grow on all amines as sources of nitrogen but still retained the ability to grow on trimethylamine as a source of carbon. Thus, unlike M. extorquens AM1 and Methylobacillus flagellatum KT, M. methylotrophus W3A1-NS does not have an additional methylamine dehydrogenase system for amine oxidation. Using a promoter-probe vector, we identified a promoter upstream of mauF and used it to construct a potential expression vector, pAYC229.
Full text
PDF







Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Chandrasekar R., Klapper M. H. Methylamine dehydrogenase and cytochrome c552 from the bacterium W3A1. J Biol Chem. 1986 Mar 15;261(8):3616–3619. [PubMed] [Google Scholar]
- Chistoserdov A. Y., Chistoserdova L. V., McIntire W. S., Lidstrom M. E. Genetic organization of the mau gene cluster in Methylobacterium extorquens AM1: complete nucleotide sequence and generation and characteristics of mau mutants. J Bacteriol. 1994 Jul;176(13):4052–4065. doi: 10.1128/jb.176.13.4052-4065.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chistoserdov A. Y., Tsygankov Y. D., Lidstrom M. E. Cloning and sequencing of the structural gene for the small subunit of methylamine dehydrogenase from Methylobacterium extorquens AM1: evidence for two tryptophan residues involved in the active center. Biochem Biophys Res Commun. 1990 Oct 15;172(1):211–216. doi: 10.1016/s0006-291x(05)80195-0. [DOI] [PubMed] [Google Scholar]
- Chistoserdov A. Y., Tsygankov Y. D., Lidstrom M. E. Genetic organization of methylamine utilization genes from Methylobacterium extorquens AM1. J Bacteriol. 1991 Sep;173(18):5901–5908. doi: 10.1128/jb.173.18.5901-5908.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ditta G., Schmidhauser T., Yakobson E., Lu P., Liang X. W., Finlay D. R., Guiney D., Helinski D. R. Plasmids related to the broad host range vector, pRK290, useful for gene cloning and for monitoring gene expression. Plasmid. 1985 Mar;13(2):149–153. doi: 10.1016/0147-619x(85)90068-x. [DOI] [PubMed] [Google Scholar]
- Eisenberg D., Schwarz E., Komaromy M., Wall R. Analysis of membrane and surface protein sequences with the hydrophobic moment plot. J Mol Biol. 1984 Oct 15;179(1):125–142. doi: 10.1016/0022-2836(84)90309-7. [DOI] [PubMed] [Google Scholar]
- Fulton G. L., Nunn D. N., Lidstrom M. E. Molecular cloning of a malyl coenzyme A lyase gene from Pseudomonas sp. strain AM1, a facultative methylotroph. J Bacteriol. 1984 Nov;160(2):718–723. doi: 10.1128/jb.160.2.718-723.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gennity J., Goldstein J., Inouye M. Signal peptide mutants of Escherichia coli. J Bioenerg Biomembr. 1990 Jun;22(3):233–269. doi: 10.1007/BF00763167. [DOI] [PubMed] [Google Scholar]
- Huitema F., van Beeumen J., van Driessche G., Duine J. A., Canters G. W. Cloning and sequencing of the gene coding for the large subunit of methylamine dehydrogenase from Thiobacillus versutus. J Bacteriol. 1993 Oct;175(19):6254–6259. doi: 10.1128/jb.175.19.6254-6259.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kenney W. C., McIntire W. Characterization of methylamine dehydrogenase from bacterium W3A1. Interaction with reductants and amino-containing compounds. Biochemistry. 1983 Aug 2;22(16):3858–3868. doi: 10.1021/bi00285a022. [DOI] [PubMed] [Google Scholar]
- McIntire W. S., Wemmer D. E., Chistoserdov A., Lidstrom M. E. A new cofactor in a prokaryotic enzyme: tryptophan tryptophylquinone as the redox prosthetic group in methylamine dehydrogenase. Science. 1991 May 10;252(5007):817–824. doi: 10.1126/science.2028257. [DOI] [PubMed] [Google Scholar]
- McIntire W. S., Weyler W. Factors affecting the production of pyrroloquinoline quinone by the methylotrophic bacterium W3A1. Appl Environ Microbiol. 1987 Sep;53(9):2183–2188. doi: 10.1128/aem.53.9.2183-2188.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meinkoth J., Wahl G. Hybridization of nucleic acids immobilized on solid supports. Anal Biochem. 1984 May 1;138(2):267–284. doi: 10.1016/0003-2697(84)90808-x. [DOI] [PubMed] [Google Scholar]
- Mohana Rao J. K., Argos P. A conformational preference parameter to predict helices in integral membrane proteins. Biochim Biophys Acta. 1986 Jan 30;869(2):197–214. doi: 10.1016/0167-4838(86)90295-5. [DOI] [PubMed] [Google Scholar]
- Owens J. D., Keddie R. M. The nitrogen nutrition of soil and herbage coryneform bacteria. J Appl Bacteriol. 1969 Sep;32(3):338–347. doi: 10.1111/j.1365-2672.1969.tb00981.x. [DOI] [PubMed] [Google Scholar]
- Ruvkun G. B., Ausubel F. M. A general method for site-directed mutagenesis in prokaryotes. Nature. 1981 Jan 1;289(5793):85–88. doi: 10.1038/289085a0. [DOI] [PubMed] [Google Scholar]
- Shirai S., Matsumoto T., Tobari J. Methylamine dehydrogenase of Pseudomonas AM1. A subunit enzyme. J Biochem. 1978 Jun;83(6):1599–1607. doi: 10.1093/oxfordjournals.jbchem.a132071. [DOI] [PubMed] [Google Scholar]
- Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
- Tsygankov Y. D., Chistoserdov A. Y. Specific-purpose broad-host-range vectors. Plasmid. 1985 Sep;14(2):118–125. doi: 10.1016/0147-619x(85)90071-x. [DOI] [PubMed] [Google Scholar]
- Ubbink M., van Kleef M. A., Kleinjan D. J., Hoitink C. W., Huitema F., Beintema J. J., Duine J. A., Canters G. W. Cloning, sequencing and expression studies of the genes encoding amicyanin and the beta-subunit of methylamine dehydrogenase from Thiobacillus versutus. Eur J Biochem. 1991 Dec 18;202(3):1003–1012. doi: 10.1111/j.1432-1033.1991.tb16462.x. [DOI] [PubMed] [Google Scholar]
- de Moura Gallo C. V., Vassetzky Y. S., Huesca M., Scherrer K. A transcription-dependent DNase I-hypersensitive site in a far upstream segment of the chicken alpha-globin gene domain coincides with a matrix attachment region. Biochem Biophys Res Commun. 1992 May 15;184(3):1226–1234. doi: 10.1016/s0006-291x(05)80013-0. [DOI] [PubMed] [Google Scholar]
- van Spanning R. J., Wansell C. W., Reijnders W. N., Oltmann L. F., Stouthamer A. H. Mutagenesis of the gene encoding amicyanin of Paracoccus denitrificans and the resultant effect on methylamine oxidation. FEBS Lett. 1990 Nov 26;275(1-2):217–220. doi: 10.1016/0014-5793(90)81475-4. [DOI] [PubMed] [Google Scholar]

