Abstract
The formate dehydrogenase-encoding fdhCAB operon and flanking genes have been cloned and sequenced from Methanobacterium thermoformicicum Z-245. fdh transcription was shown to be initiated 21 bp upstream from fdhC, although most fdh transcripts terminated or were processed between fdhC and fdhA. The resulting fdhC, fdhAB, and fdhCAB transcripts were present at all growth stages in cells growing on formate but were barely detectable during early exponential growth on H2 plus CO2. The levels of the fdh transcripts did, however, increase dramatically in cells growing on H2 plus CO2, coincident with the decrease in the growth rate and the onset of constant methanogenesis that occurred when culture densities reached an optical density at 600 nm of approximately 0.5. The mth transcript that encodes the H2-dependent methenyl-H4 MPT reductase (MTH) and the frh and mvh transcripts that encode the coenzyme F420-reducing (FRH) and nonreducing (MVH) hydrogenases, respectively, were also present in cells growing on formate, consistent with the synthesis of three hydrogenases, MTH, FRH, and MVH, in the absence of exogenously supplied H2. Reducing the H2 supply to M. thermoformicicum cells growing on H2 plus CO2 reduced the growth rate and CH4 production but increased frh and fdh transcription and also increased transcription of the mtd, mer, and mcr genes that encode enzymes that catalyze steps 4, 5, and 7, respectively, in the pathway of CO2 reduction to CH4. Reducing the H2 supply to a level insufficient for growth resulted in the disappearance of all methane gene transcripts except the mcr transcript, which increased. Regions flanking the fdhCAB operon in M. thermoformicicum Z-245 were used as probes to clone the homologous region from the Methanobacterium thermoautotrophicum deltaH genome. Sequencing revealed the presence of very similar genes except that the genome of M. thermoautotrophicum, a methanogen incapable of growth on formate, lacked the fdhCAB operon.
Full Text
The Full Text of this article is available as a PDF (6.7 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Alex L. A., Reeve J. N., Orme-Johnson W. H., Walsh C. T. Cloning, sequence determination, and expression of the genes encoding the subunits of the nickel-containing 8-hydroxy-5-deazaflavin reducing hydrogenase from Methanobacterium thermoautotrophicum delta H. Biochemistry. 1990 Aug 7;29(31):7237–7244. doi: 10.1021/bi00483a011. [DOI] [PubMed] [Google Scholar]
- Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
- Baron S. F., Ferry J. G. Reconstitution and properties of a coenzyme F420-mediated formate hydrogenlyase system in Methanobacterium formicicum. J Bacteriol. 1989 Jul;171(7):3854–3859. doi: 10.1128/jb.171.7.3854-3859.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bonacker L. G., Baudner S., Thauer R. K. Differential expression of the two methyl-coenzyme M reductases in Methanobacterium thermoautotrophicum as determined immunochemically via isoenzyme-specific antisera. Eur J Biochem. 1992 May 15;206(1):87–92. doi: 10.1111/j.1432-1033.1992.tb16904.x. [DOI] [PubMed] [Google Scholar]
- Brown J. W., Daniels C. J., Reeve J. N. Gene structure, organization, and expression in archaebacteria. Crit Rev Microbiol. 1989;16(4):287–338. doi: 10.3109/10408418909105479. [DOI] [PubMed] [Google Scholar]
- Cao X. J., Krzycki J. A. Acetate-dependent methylation of two corrinoid proteins in extracts of Methanosarcina barkeri. J Bacteriol. 1991 Sep;173(17):5439–5448. doi: 10.1128/jb.173.17.5439-5448.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fukuzawa H., Suzuki E., Komukai Y., Miyachi S. A gene homologous to chloroplast carbonic anhydrase (icfA) is essential to photosynthetic carbon dioxide fixation by Synechococcus PCC7942. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4437–4441. doi: 10.1073/pnas.89.10.4437. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hausner W., Frey G., Thomm M. Control regions of an archaeal gene. A TATA box and an initiator element promote cell-free transcription of the tRNA(Val) gene of Methanococcus vannielii. J Mol Biol. 1991 Dec 5;222(3):495–508. doi: 10.1016/0022-2836(91)90492-o. [DOI] [PubMed] [Google Scholar]
- Hochheimer A., Schmitz R. A., Thauer R. K., Hedderich R. The tungsten formylmethanofuran dehydrogenase from Methanobacterium thermoautotrophicum contains sequence motifs characteristic for enzymes containing molybdopterin dinucleotide. Eur J Biochem. 1995 Dec 15;234(3):910–920. doi: 10.1111/j.1432-1033.1995.910_a.x. [DOI] [PubMed] [Google Scholar]
- Jayaratne P., Bronner D., MacLachlan P. R., Dodgson C., Kido N., Whitfield C. Cloning and analysis of duplicated rfbM and rfbK genes involved in the formation of GDP-mannose in Escherichia coli O9:K30 and participation of rfb genes in the synthesis of the group I K30 capsular polysaccharide. J Bacteriol. 1994 Jun;176(11):3126–3139. doi: 10.1128/jb.176.11.3126-3139.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jones W. J., Nagle D. P., Jr, Whitman W. B. Methanogens and the diversity of archaebacteria. Microbiol Rev. 1987 Mar;51(1):135–177. doi: 10.1128/mr.51.1.135-177.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kletzin A. Molecular characterisation of a DNA ligase gene of the extremely thermophilic archaeon Desulfurolobus ambivalens shows close phylogenetic relationship to eukaryotic ligases. Nucleic Acids Res. 1992 Oct 25;20(20):5389–5396. doi: 10.1093/nar/20.20.5389. [DOI] [PMC free article] [PubMed] [Google Scholar]
- May H. D., Patel P. S., Ferry J. G. Effect of molybdenum and tungsten on synthesis and composition of formate dehydrogenase in Methanobacterium formicicum. J Bacteriol. 1988 Aug;170(8):3384–3389. doi: 10.1128/jb.170.8.3384-3389.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meese E., Olson S., Leis L., Trent J. Quick method for high yields of lambda bacteriophage DNA. Nucleic Acids Res. 1990 Apr 11;18(7):1923–1923. doi: 10.1093/nar/18.7.1923. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morgan R. M., Pihl T. D., Nölling J., Reeve J. N. Hydrogen regulation of growth, growth yields, and methane gene transcription in Methanobacterium thermoautotrophicum deltaH. J Bacteriol. 1997 Feb;179(3):889–898. doi: 10.1128/jb.179.3.889-898.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mukhopadhyay B., Purwantini E., Pihl T. D., Reeve J. N., Daniels L. Cloning, sequencing, and transcriptional analysis of the coenzyme F420-dependent methylene-5,6,7,8-tetrahydromethanopterin dehydrogenase gene from Methanobacterium thermoautotrophicum strain Marburg and functional expression in Escherichia coli. J Biol Chem. 1995 Feb 10;270(6):2827–2832. doi: 10.1074/jbc.270.6.2827. [DOI] [PubMed] [Google Scholar]
- Nölling J., Pihl T. D., Reeve J. N. Cloning, sequencing, and growth phase-dependent transcription of the coenzyme F420-dependent N5,N10-methylenetetrahydromethanopterin reductase-encoding genes from Methanobacterium thermoautotrophicum delta H and Methanopyrus kandleri. J Bacteriol. 1995 Dec;177(24):7238–7244. doi: 10.1128/jb.177.24.7238-7244.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nölling J., Pihl T. D., Vriesema A., Reeve J. N. Organization and growth phase-dependent transcription of methane genes in two regions of the Methanobacterium thermoautotrophicum genome. J Bacteriol. 1995 May;177(9):2460–2468. doi: 10.1128/jb.177.9.2460-2468.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Patel P. S., Ferry J. G. Characterization of the upstream region of the formate dehydrogenase operon of Methanobacterium formicicum. J Bacteriol. 1988 Aug;170(8):3390–3395. doi: 10.1128/jb.170.8.3390-3395.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pihl T. D., Sharma S., Reeve J. N. Growth phase-dependent transcription of the genes that encode the two methyl coenzyme M reductase isoenzymes and N5-methyltetrahydromethanopterin:coenzyme M methyltransferase in Methanobacterium thermoautotrophicum delta H. J Bacteriol. 1994 Oct;176(20):6384–6391. doi: 10.1128/jb.176.20.6384-6391.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reeve J. N., Beckler G. S., Cram D. S., Hamilton P. T., Brown J. W., Krzycki J. A., Kolodziej A. F., Alex L., Orme-Johnson W. H., Walsh C. T. A hydrogenase-linked gene in Methanobacterium thermoautotrophicum strain delta H encodes a polyferredoxin. Proc Natl Acad Sci U S A. 1989 May;86(9):3031–3035. doi: 10.1073/pnas.86.9.3031. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rivers S. L., McNairn E., Blasco F., Giordano G., Boxer D. H. Molecular genetic analysis of the moa operon of Escherichia coli K-12 required for molybdenum cofactor biosynthesis. Mol Microbiol. 1993 Jun;8(6):1071–1081. doi: 10.1111/j.1365-2958.1993.tb01652.x. [DOI] [PubMed] [Google Scholar]
- Robison K., Gilbert W., Church G. M. Large scale bacterial gene discovery by similarity search. Nat Genet. 1994 Jun;7(2):205–214. doi: 10.1038/ng0694-205. [DOI] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schauer N. L., Ferry J. G. Metabolism of formate in Methanobacterium formicicum. J Bacteriol. 1980 Jun;142(3):800–807. doi: 10.1128/jb.142.3.800-807.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schauer N. L., Ferry J. G. Properties of formate dehydrogenase in Methanobacterium formicicum. J Bacteriol. 1982 Apr;150(1):1–7. doi: 10.1128/jb.150.1.1-7.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shuber A. P., Orr E. C., Recny M. A., Schendel P. F., May H. D., Schauer N. L., Ferry J. G. Cloning, expression, and nucleotide sequence of the formate dehydrogenase genes from Methanobacterium formicicum. J Biol Chem. 1986 Oct 5;261(28):12942–12947. [PubMed] [Google Scholar]
- Sparling R., Daniels L. Regulation of formate dehydrogenase activity in Methanococcus thermolithotrophicus. J Bacteriol. 1990 Mar;172(3):1464–1469. doi: 10.1128/jb.172.3.1464-1469.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Touzel J. P., Conway de Macario E., Nölling J., De Vos W. M., Zhilina T., Lysenko A. M. DNA relatedness among some thermophilic members of the genus Methanobacterium: emendation of the species Methanobacterium thermoautotrophicum and rejection of Methanobacterium thermoformicicum as a synonym of Methanobacterium thermoautotrophicum. Int J Syst Bacteriol. 1992 Jul;42(3):408–411. doi: 10.1099/00207713-42-3-408. [DOI] [PubMed] [Google Scholar]
- Vermeij P., van der Steen R. J., Keltjens J. T., Vogels G. D., Leisinger T. Coenzyme F390 synthetase from Methanobacterium thermoautotrophicum Marburg belongs to the superfamily of adenylate-forming enzymes. J Bacteriol. 1996 Jan;178(2):505–510. doi: 10.1128/jb.178.2.505-510.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weil C. F., Cram D. S., Sherf B. A., Reeve J. N. Structure and comparative analysis of the genes encoding component C of methyl coenzyme M reductase in the extremely thermophilic archaebacterium Methanothermus fervidus. J Bacteriol. 1988 Oct;170(10):4718–4726. doi: 10.1128/jb.170.10.4718-4726.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- White W. B., Ferry J. G. Identification of formate dehydrogenase-specific mRNA species and nucleotide sequence of the fdhC gene of Methanobacterium formicicum. J Bacteriol. 1992 Aug;174(15):4997–5004. doi: 10.1128/jb.174.15.4997-5004.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
- Yie Y., Wei Z., Tien P. A simplified and reliable protocol for plasmid DNA sequencing: fast miniprep and denaturation. Nucleic Acids Res. 1993 Jan 25;21(2):361–361. doi: 10.1093/nar/21.2.361. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zirngibl C., Van Dongen W., Schwörer B., Von Bünau R., Richter M., Klein A., Thauer R. K. H2-forming methylenetetrahydromethanopterin dehydrogenase, a novel type of hydrogenase without iron-sulfur clusters in methanogenic archaea. Eur J Biochem. 1992 Sep 1;208(2):511–520. doi: 10.1111/j.1432-1033.1992.tb17215.x. [DOI] [PubMed] [Google Scholar]