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Random mutagenesis identifies factors involved in formate-dependent growth of the methanogenic archaeon Methanococcus maripaludis

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Abstract

Methane is a key intermediate in the carbon cycle and biologically produced by methanogenic archaea. Most methanogens are able to conserve energy by reducing CO2 to methane using molecular hydrogen as electron donor (hydrogenotrophic methanogenesis), but several hydrogenotrophic methanogens can also use formate as electron donor for methanogenesis. Formate dehydrogenase (Fdh) oxidizes formate to CO2 and is involved in funneling reducing equivalents into the methanogenic pathway, but details on other factors relevant for formate-dependent physiology of methanogens are not available. To learn more about the factors involved in formate-dependent growth of Methanococcus maripaludis strain JJ, we used a recently developed system for random in vitro mutagenesis, which is based on a modified insect transposable element to create 2,865 chromosomal transposon mutants and screened them for impaired growth on formate. Of 12 M. maripaludis transposon-induced mutants exhibiting this phenotype, the transposon insertion sites in the chromosome were mapped. Among the genes, apparently affecting formate-dependent growth were those encoding archaeal transcription factor S, a regulator of ion transport, and carbon monoxide dehydrogenase/acetyl-CoA synthase. Interestingly, in seven of the mutants, transposons were localized in a 10.2 kb region where Fdh1, one of two Fdh isoforms in the organism, is encoded. Two transcription start sites within the 10.2 kb region could be mapped, and quantification of transcripts revealed that transposon insertion in this region diminished fdhA1 expression due to polar effects.

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Acknowledgments

We thank Deniz Seyhan (Frankfurt am Main, Germany) for his help in screening mini-MAR367 mutants and Marina Totrova for excellent technical assistance. The authors declare that they have no conflict of interest. This work was supported by a grant from the Deutsche Forschungsgemeinschaft (via SFB 902) to M.R.

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Authors and Affiliations

  1. Institut für Mikrobiologie, Technische Universität Dresden, 01062, Dresden, Germany

    Christian Sattler, Sandro Wolf, Julia Fersch & Michael Rother

  2. Institut für Molekulare Biowissenschaften, Johann Wolfgang Goethe-Universität Frankfurt, Frankfurt, Germany

    Christian Sattler, Stefan Goetz & Michael Rother

Authors
  1. Christian Sattler
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  2. Sandro Wolf
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  3. Julia Fersch
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  4. Stefan Goetz
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  5. Michael Rother
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Corresponding author

Correspondence to Michael Rother.

Additional information

Communicated by G. Klug.

Nucleotide sequence data reported are available in the GenBank database under accession number KC342234.

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Sattler, C., Wolf, S., Fersch, J. et al. Random mutagenesis identifies factors involved in formate-dependent growth of the methanogenic archaeon Methanococcus maripaludis . Mol Genet Genomics 288, 413–424 (2013). https://doi.org/10.1007/s00438-013-0756-6

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  • DOI: https://doi.org/10.1007/s00438-013-0756-6

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