J. Bacteriol., Dec 1996, 6849-6856, Vol 178, No. 23
Copyright © 1996, American Society for Microbiology

Analysis of the CO dehydrogenase/acetyl-coenzyme A synthase operon of Methanosarcina thermophila

JA Maupin-Furlow and JG Ferry
Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park 16802-4500, USA.

The cdhABC genes encoding the respective alpha, epsilon, and beta subunits of the five-subunit (alpha, beta, gamma, delta, and epsilon) CO dehydrogenase/acetyl-coenzyme synthase (CODH/ACS) complex from Methanosarcina thermophila were cloned and sequenced. Northern (RNA) blot analyses indicated that the cdh genes encoding the five subunits and an open reading frame (ORF1) with unknown function are cotranscribed during growth on acetate. Northern blot and primer extension analyses suggested that mRNA processing and multiple promoters may be involved in cdh transcript synthesis. The putative CdhA (alpha subunit) and CdhB (epsilon subunit) proteins each have 40% identity to CdhA and CdhB of the CODH/ACS complex from Methanosaeta soehngenii. The cdhC gene encodes the beta subunit (CdhC) of the CODH/ACS complex from M. thermophila. The N-terminal 397 amino acids of CdhC are 42% identical to the C-terminal half of the alpha subunit of CODH/ACS from the acetogenic anaerobe Clostridium thermoaceticum. Sequence analysis suggested potential structures and functions for the previously uncharacterized beta subunit from M. thermophila. The deduced protein sequence of ORF1, located between the cdhC and cdhD genes, has 29% identity to NifH2 from Methanobacterium ivanovii.

This article has been cited by other articles:

• Lessner, D. J., Li, L., Li, Q., Rejtar, T., Andreev, V. P., Reichlen, M., Hill, K., Moran, J. J., Karger, B. L., Ferry, J. G. (2006). An unconventional pathway for reduction of CO2 to methane in CO-grown Methanosarcina acetivorans revealed by proteomics. Proc. Natl. Acad. Sci. USA 103: 17921-17926 [Abstract] [Full Text]  
• Apolinario, E. E., Jackson, K. M., Sowers, K. R. (2005). Development of a Plasmid-Mediated Reporter System for In Vivo Monitoring of Gene Expression in the Archaeon Methanosarcina acetivorans. Appl. Environ. Microbiol. 71: 4914-4918 [Abstract] [Full Text]  
• Hendrickson, E. L., Kaul, R., Zhou, Y., Bovee, D., Chapman, P., Chung, J., Conway de Macario, E., Dodsworth, J. A., Gillett, W., Graham, D. E., Hackett, M., Haydock, A. K., Kang, A., Land, M. L., Levy, R., Lie, T. J., Major, T. A., Moore, B. C., Porat, I., Palmeiri, A., Rouse, G., Saenphimmachak, C., Soll, D., Van Dien, S., Wang, T., Whitman, W. B., Xia, Q., Zhang, Y., Larimer, F. W., Olson, M. V., Leigh, J. A. (2004). Complete Genome Sequence of the Genetically Tractable Hydrogenotrophic Methanogen Methanococcus maripaludis. J. Bacteriol. 186: 6956-6969 [Abstract] [Full Text]  
• Gencic, S., Grahame, D. A. (2003). Nickel in Subunit beta of the Acetyl-CoA Decarbonylase/Synthase Multienzyme Complex in Methanogens. CATALYTIC PROPERTIES AND EVIDENCE FOR A BINUCLEAR Ni-Ni SITE. J. Biol. Chem. 278: 6101-6110 [Abstract] [Full Text]  
• Thomas, N. A., Jarrell, K. F. (2001). Characterization of Flagellum Gene Families of Methanogenic Archaea and Localization of Novel Flagellum Accessory Proteins. J. Bacteriol. 183: 7154-7164 [Abstract] [Full Text]  
• Jeon, W. B., Cheng, J., Ludden, P. W. (2001). Purification and Characterization of Membrane-associated CooC Protein and Its Functional Role in the Insertion of Nickel into Carbon Monoxide Dehydrogenase from Rhodospirillum rubrum. J. Biol. Chem. 276: 38602-38609 [Abstract] [Full Text]  
• Loke, H.-K., Bennett, G. N., Lindahl, P. A. (2000). Active acetyl-CoA synthase from Clostridium thermoaceticum obtained by cloning and heterologous expression of acsAB in Escherichia coli. Proc. Natl. Acad. Sci. USA 10.1073/pnas.220404397v1 [Abstract] [Full Text]  
• Leartsakulpanich, U., Antonkine, M. L., Ferry, J. G. (2000). Site-Specific Mutational Analysis of a Novel Cysteine Motif Proposed To Ligate the 4Fe-4S Cluster in the Iron-Sulfur Flavoprotein of the Thermophilic Methanoarchaeon Methanosarcina thermophila. J. Bacteriol. 182: 5309-5316 [Abstract] [Full Text]  
• Chien, Y.-T., Auerbuch, V., Brabban, A. D., Zinder, S. H. (2000). Analysis of Genes Encoding an Alternative Nitrogenase in the Archaeon Methanosarcina barkeri 227. J. Bacteriol. 182: 3247-3253 [Abstract] [Full Text]  
• Murakami, E., Ragsdale, S. W. (2000). Evidence for Intersubunit Communication during Acetyl-CoA Cleavage by the Multienzyme CO Dehydrogenase/Acetyl-CoA Synthase Complex from Methanosarcina thermophila. EVIDENCE THAT THE beta SUBUNIT CATALYZES C-C AND C-S BOND CLEAVAGE. J. Biol. Chem. 275: 4699-4707 [Abstract] [Full Text]  
• Becker, D. F., Leartsakulpanich, U., Surerus, K. K., Ferry, J. G., Ragsdale, S. W. (1998). Electrochemical and Spectroscopic Properties of the Iron-Sulfur Flavoprotein from Methanosarcina thermophila. J. Biol. Chem. 273: 26462-26469 [Abstract] [Full Text]  
• Burke, S. A., Lo, S. L., Krzycki, J. A. (1998). Clustered Genes Encoding the Methyltransferases of Methanogenesis from Monomethylamine. J. Bacteriol. 180: 3432-3440 [Abstract] [Full Text]  
• Kessler, P. S., Blank, C., Leigh, J. A. (1998). The nif Gene Operon of the Methanogenic Archaeon Methanococcus maripaludis. J. Bacteriol. 180: 1504-1511 [Abstract] [Full Text]  
• Spangler, N. J., Meyers, M. R., Gierke, K. L., Kerby, R. L., Roberts, G. P., Ludden, P. W. (1998). Substitution of Valine for Histidine 265 in Carbon Monoxide Dehydrogenase from Rhodospirillum rubrum Affects Activity and Spectroscopic States. J. Biol. Chem. 273: 4059-4064 [Abstract] [Full Text]  
• Loke, H.-K., Bennett, G. N., Lindahl, P. A. (2000). Active acetyl-CoA synthase from Clostridium thermoaceticum obtained by cloning and heterologous expression of acsAB in Escherichia coli. Proc. Natl. Acad. Sci. USA 97: 12530-12535 [Abstract] [Full Text]