The genome of M. acetivorans reveals extensive metabolic and physiological diversity.

Methanogenesis, the biological production of methane, plays a pivotal role in the global carbon cycle and contributes significantly to global warming. The majority of methane in nature is derived from acetate. Here we report the complete genome sequence of an acetate-utilizing methanogen, Methanosarcina acetivorans C2A. Methanosarcineae are the most metabolically diverse methanogens, thrive in a broad range of environments, and are unique among the Archaea in forming complex multicellular structures. This diversity is reflected in the genome of M. acetivorans. At 5,751,492 base pairs it is by far the largest known archaeal genome. The 4524 open reading frames code for a strikingly wide and unanticipated variety of metabolic and cellular capabilities. The presence of novel methyltransferases indicates the likelihood of undiscovered natural energy sources for methanogenesis, whereas the presence of single-subunit carbon monoxide dehydrogenases raises the possibility of nonmethanogenic growth. Although motility has not been observed in any Methanosarcineae, a flagellin gene cluster and two complete chemotaxis gene clusters were identified. The availability of genetic methods, coupled with its physiological and metabolic diversity, makes M. acetivorans a powerful model organism for the study of archaeal biology. [Sequence, data, annotations and analyses are available at http://www-genome.wi.mit.edu/.]

[1]  Y. Le Loir,et al.  Respiration Capacity of the Fermenting BacteriumLactococcus lactis and Its Positive Effects on Growth and Survival , 2001, Journal of bacteriology.

[2]  T. Ficht,et al.  Interruption of the cydB Locus inBrucella abortus Attenuates Intracellular Survival and Virulence in the Mouse Model of Infection , 2001, Journal of bacteriology.

[3]  S. Salzberg,et al.  Optimized multiplex PCR: efficiently closing a whole-genome shotgun sequencing project. , 1999, Genomics.

[4]  E. C. de Macario,et al.  Immunochemical differences among Methanosarcina mazei S-6 morphologic forms , 1992, Journal of bacteriology.

[5]  Z. Kelman The replication origin of archaea is finally revealed. , 2000, Trends in biochemical sciences.

[6]  K. Jarrell,et al.  Effect of changes in mineral composition and growth temperature on filament length and flagellation in the Archaeon Methanospirillum hungatei , 1993, Archives of Microbiology.

[7]  J. Vorholt,et al.  An Escherichia coli hydrogenase-3-type hydrogenase in methanogenic archaea. , 1998, European journal of biochemistry.

[8]  C. James,et al.  The Amber Codon in the Gene Encoding the Monomethylamine Methyltransferase Isolated from Methanosarcina barkeri Is Translated as a Sense Codon* , 2001, The Journal of Biological Chemistry.

[9]  S. Schuster,et al.  The fla gene cluster is involved in the biogenesis of flagella in Halobacterium salinarum , 2001, Molecular microbiology.

[10]  Sean R. Eddy,et al.  Biological Sequence Analysis: Probabilistic Models of Proteins and Nucleic Acids , 1998 .

[11]  E. Conway de Macario,et al.  Integration of foreign DNA in an intergenic region of the archaeon Methanosarcina mazei without effect on transcription of adjacent genes. , 1996, Journal of molecular biology.

[12]  N. Baliga,et al.  Is gene expression in Halobacterium NRC‐1 regulated by multiple TBP and TFB transcription factors? , 2000, Molecular microbiology.

[13]  S. Salzberg,et al.  Improved microbial gene identification with GLIMMER. , 1999, Nucleic acids research.

[14]  K. Sowers,et al.  A genetic system for Archaea of the genus Methanosarcina: liposome-mediated transformation and construction of shuttle vectors. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[15]  B. Rost,et al.  Topology prediction for helical transmembrane proteins at 86% accuracy–Topology prediction at 86% accuracy , 1996, Protein science : a publication of the Protein Society.

[16]  J. Krzycki,et al.  The Trimethylamine Methyltransferase Gene and Multiple Dimethylamine Methyltransferase Genes of Methanosarcina barkeri Contain In-Frame and Read-Through Amber Codons , 2000, Journal of bacteriology.

[17]  Fan Yang,et al.  TIGRFAMs: a protein family resource for the functional identification of proteins , 2001, Nucleic Acids Res..

[18]  J. Zeikus,et al.  Association of hydrogen metabolism with unitrophic or mixotrophic growth of Methanosarcina barkeri on carbon monoxide , 1984, Journal of bacteriology.

[19]  J. Krzycki,et al.  Sequence and transcript analysis of a novel Methanosarcina barkeri methyltransferase II homolog and its associated corrinoid protein homologous to methionine synthase , 1996, Journal of bacteriology.

[20]  J. Takagi,et al.  A Novel Ca2+ Binding β Hairpin Loop Better Resembles Integrin Sequence Motifs Than the EF Hand , 2000, Cell.

[21]  D. Söll,et al.  Context-dependent anticodon recognition by class I lysyl-tRNA synthetases. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[22]  H. König,et al.  Uridine and dolichyl diphosphate activated oligosaccharides are intermediates in the biosynthesis of the S-layer glycoprotein of Methanothermus fervidus , 1989, Archives of Microbiology.

[23]  O. Kandler,et al.  Cell wall polymers in Archaea (Archaebacteria) , 1998, Cellular and Molecular Life Sciences CMLS.

[24]  R. Gunsalus,et al.  Disaggregation of Methanosarcina spp. and Growth as Single Cells at Elevated Osmolarity , 1993, Applied and environmental microbiology.

[25]  R. Hedderich,et al.  Purification and catalytic properties of Ech hydrogenase from Methanosarcina barkeri. , 1999, European journal of biochemistry.

[26]  K. Jarrell,et al.  Cloning and sequencing of a multigene family encoding the flagellins of Methanococcus voltae , 1991, Journal of bacteriology.

[27]  K. Nagahisa,et al.  Sequence and transcriptional studies of five clustered flagellin genes from hyperthermophilic archaeon Pyrococcus kodakaraensis KOD1. , 1999, FEMS microbiology letters.

[28]  A Grigoriev,et al.  Analyzing genomes with cumulative skew diagrams. , 1998, Nucleic acids research.

[29]  J E Sulston,et al.  Short-insert libraries as a method of problem solving in genome sequencing. , 1998, Genome research.

[30]  Steven W. Leavit Biogeochemistry, An Analysis of Global Change , 1998 .

[31]  W. Metcalf,et al.  An Anaerobic, Intrachamber Incubator for Growth ofMethanosarcina spp. on Methanol-Containing Solid Media , 1998, Applied and Environmental Microbiology.

[32]  E. Conway de Macario,et al.  The molecular chaperone system and other anti-stress mechanisms in archaea. , 2001, Frontiers in bioscience : a journal and virtual library.

[33]  P. Boccazzi,et al.  Directed Mutagenesis and Plasmid-Based Complementation in the Methanogenic Archaeon Methanosarcina acetivorans C2A Demonstrated by Genetic Analysis of Proline Biosynthesis , 2002, Journal of bacteriology.

[34]  C. Ponting,et al.  PAS: a multifunctional domain family comes to light , 1997, Current Biology.

[35]  W. Metcalf,et al.  Molecular, genetic, and biochemical characterization of the serC gene of Methanosarcina barkeri Fusaro , 1996, Journal of bacteriology.

[36]  R. Thauer,et al.  Two F420-reducing hydrogenases in Methanosarcina barkeri , 1998, Archives of Microbiology.

[37]  K. Sowers,et al.  Isolation and Characterization of a Methylotrophic Marine Methanogen, Methanococcoides methylutens gen. nov., sp. nov , 1983, Applied and environmental microbiology.

[38]  O. Kandler,et al.  Chapter 8 Cell envelopes of archaea: Structure and chemistry , 1993 .

[39]  R S Wolfe,et al.  A novel pH2 control on the expression of flagella in the hyperthermophilic strictly hydrogenotrophic methanarchaeaon Methanococcus jannaschii. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[40]  S. Bell,et al.  Mechanism and regulation of transcription in archaea. , 2001, Current opinion in microbiology.

[41]  S. F. Baron,et al.  Methanosarcina acetivorans sp. nov., an Acetotrophic Methane-Producing Bacterium Isolated from Marine Sediments , 1984, Applied and environmental microbiology.

[42]  E. Conway de Macario,et al.  Conservation and variability in Archaea: protein antigens with tandem repeats encoded by a cluster of genes with common motifs in Methanosarcina mazei S-6. , 1995, Gene.

[43]  K. Jarrell,et al.  Posttranslational Processing of Methanococcus voltae Preflagellin by Preflagellin Peptidases of M. voltae and Other Methanogens , 2000, Journal of bacteriology.

[44]  Dorothea K. Thompson,et al.  Expression and heat‐responsive regulation of a TFIIB homologue from the archaeon Haloferax volcanii , 1999, Molecular microbiology.

[45]  R. Gunsalus,et al.  Aerobic regulation of cytochrome d oxidase (cydAB ) operon expression in Escherichia coli: roles of Fnr and ArcA in repression and activation , 1997, Molecular microbiology.

[46]  T. Thiel Characterization of genes for an alternative nitrogenase in the cyanobacterium Anabaena variabilis , 1993, Journal of bacteriology.

[47]  Stephen H. Zinder,et al.  Physiological Ecology of Methanogens , 1993 .

[48]  M. Pritchett,et al.  In vivo transposon mutagenesis of the methanogenic archaeon Methanosarcina acetivorans C2A using a modified version of the insect mariner-family transposable element Himar1. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[49]  P. Boccazzi,et al.  Generation of Dominant Selectable Markers for Resistance to Pseudomonic Acid by Cloning and Mutagenesis of theileS Gene from the Archaeon Methanosarcina barkeri Fusaro , 2000, Journal of bacteriology.

[50]  S. Kay,et al.  Photoactive yellow protein: a structural prototype for the three-dimensional fold of the PAS domain superfamily. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[51]  V. Thorsson,et al.  Genome sequence of Halobacterium species NRC-1. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[52]  F. Robb,et al.  Complete sequence and gene organization of the genome of a hyper-thermophilic archaebacterium, Pyrococcus horikoshii OT3. , 1998, DNA research : an international journal for rapid publication of reports on genes and genomes.