Role of multiple gene copies in particulate methane monooxygenase activity in the methane-oxidizing bacterium Methylococcus capsulatus Bath

Sergei Stolyar, Andria Costello Staniec, Tonya L. Peeples, Mary E. Lidstrom

Research output: Contribution to journalArticle

95 Citations (Scopus)

Abstract

Genes for the subunits of particulate methane monooxygenase, PmoABC, have been sequenced from the γ-proteobacterial methanotroph Methylococcus capsulatus Bath. M. capsulatus Bath contains two complete copies of pmoCAB, as well as a third copy of pmoC. The two pmoCAB regions were almost identical at the nucleotide sequence level, differing in only 13 positions in 3183 bp. At the amino acid level, each translated gene product contained only one differing residue in each copy. However, the pmoC3 sequence was more divergent from the two other pmoC copies at both the far N-terminus and far C-terminus. Chromosomal insertion mutations were generated in all seven genes. Null mutants could not be obtained for pmoC3, suggesting that it may play an essential role in growth on methane. Null mutants were obtained for pmoC1, pmoC2, pmoA1, pmoA2, pmoB1 and pmoB2. All of these mutants grew on methane, demonstrating that both gene copies were functional. Copy 1 mutants showed about two-thirds of the wild-type whole-cell methane oxidation rate, while copy 2 mutants showed only about one-third of the wild-type rate, indicating that both gene copies were necessary for wild-type particulate methane monooxygenase activity. It was not possible to obtain double null mutants that were defective in both pmo copies, which may indicate that some expression of pMMO is important for growth.

Original languageEnglish (US)
Pages (from-to)1235-1244
Number of pages10
JournalMicrobiology
Volume145
Issue number5
StatePublished - May 1999
Externally publishedYes

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methane monooxygenase
Methylococcus capsulatus
Methylococcaceae
Baths
Methane
Genes
Insertional Mutagenesis
Growth
Amino Acids

Keywords

  • Duplicate genes
  • Methane monooxygenase
  • Methanotroph
  • pmo

ASJC Scopus subject areas

  • Microbiology

Cite this

Role of multiple gene copies in particulate methane monooxygenase activity in the methane-oxidizing bacterium Methylococcus capsulatus Bath. / Stolyar, Sergei; Staniec, Andria Costello; Peeples, Tonya L.; Lidstrom, Mary E.

In: Microbiology, Vol. 145, No. 5, 05.1999, p. 1235-1244.

Research output: Contribution to journalArticle

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AB - Genes for the subunits of particulate methane monooxygenase, PmoABC, have been sequenced from the γ-proteobacterial methanotroph Methylococcus capsulatus Bath. M. capsulatus Bath contains two complete copies of pmoCAB, as well as a third copy of pmoC. The two pmoCAB regions were almost identical at the nucleotide sequence level, differing in only 13 positions in 3183 bp. At the amino acid level, each translated gene product contained only one differing residue in each copy. However, the pmoC3 sequence was more divergent from the two other pmoC copies at both the far N-terminus and far C-terminus. Chromosomal insertion mutations were generated in all seven genes. Null mutants could not be obtained for pmoC3, suggesting that it may play an essential role in growth on methane. Null mutants were obtained for pmoC1, pmoC2, pmoA1, pmoA2, pmoB1 and pmoB2. All of these mutants grew on methane, demonstrating that both gene copies were functional. Copy 1 mutants showed about two-thirds of the wild-type whole-cell methane oxidation rate, while copy 2 mutants showed only about one-third of the wild-type rate, indicating that both gene copies were necessary for wild-type particulate methane monooxygenase activity. It was not possible to obtain double null mutants that were defective in both pmo copies, which may indicate that some expression of pMMO is important for growth.

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