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Genetic analysis of MA4079, an aldehyde dehydrogenase homolog, in Methanosarcina acetivorans

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Abstract

When Methanosarcina acetivorans grows on carbon monoxide (CO), it synthesizes high levels of a protein, MA4079, homologous to aldehyde dehydrogenases. To investigate the role of MA4079 in M. acetivorans, mutants lacking the encoding gene were generated and phenotypically analyzed. Loss of MA4079 had no effect on methylotrophic growth but led to complete abrogation of methylotrophic growth in the presence of even small amounts of CO, which indicated the mutant’s inability to acclimate to the presence of this toxic gas. Prolonged incubation with CO allowed the isolation of a strain in which the effect of MA4079 deletion is suppressed. The strain, designated Mu3, tolerated the presence of high CO partial pressures even better than the wild type. Immunological analysis using antisera against MA4079 suggested that it is not abundant in M. acetivorans. Comparison of proteins differentially abundant in Mu3 and the wild type revealed an elevated level of methyl-coenzyme M reductase and a decreased level of one isoform of carbon monoxide dehydrogenase/acetyl-coenzyme A synthase, which suggests that pleiotropic mutation(s) compensating for the loss of MA4079 affected catabolism. The data presented point toward a role of MA4079 to enable M. acetivorans to properly acclimate to CO.

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Acknowledgments

V. Müller, Goethe-Universität Frankfurt, and H. Michel, Max-Planck-Institut für Biophysik, are gratefully acknowledged for their support. This work was supported by a grant from the Deutsche Forschungsgemeinschaft (RO 2445/6-1) to M.R. and by grants from the Bundesministerium für Bildung und Forschung and the European Union ESFRI Instruct initiative (Membrane Protein Core Center) and by the Max-Planck Gesellschaft.

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  1. Ellen Oelgeschläger

    Present address: Bayer Healthcare, Chempark Gebäude E39, 51368, Leverkusen, Germany

Authors and Affiliations

  1. Institut für Molekulare Biowissenschaften, Molekulare Mikrobiologie und Bioenergetik, Johann Wolfgang Goethe-Universität Frankfurt, Max-von-Laue-Str. 9, 60438, Frankfurt am Main, Germany

    Michael Kliefoth, Nicole Matschiavelli, Ellen Oelgeschläger & Michael Rother

  2. Molekulare Membranbiologie, Max-Planck-Institut für Biophysik, Frankfurt am Main, Germany

    Julian D. Langer

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Correspondence to Michael Rother.

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Communicated by Erko Stackebrandt.

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Kliefoth, M., Langer, J.D., Matschiavelli, N. et al. Genetic analysis of MA4079, an aldehyde dehydrogenase homolog, in Methanosarcina acetivorans . Arch Microbiol 194, 75–85 (2012). https://doi.org/10.1007/s00203-011-0727-4

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