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Title: Posttranslational Methylation of Arginine in Methyl Coenzyme M Reductase Has a Profound Impact on both Methanogenesis and Growth of Methanococcus maripaludis

Abstract

Catalyzing the key step for anaerobic production and/or oxidation of methane and likely other short-chain alkanes, methyl coenzyme M reductase (Mcr) and its homologs play a key role in the global carbon cycle. The McrA subunit possesses up to five conserved posttranslational modifications (PTMs) at its active site. It was previously suggested that methanogenesis marker protein 10 (Mmp10) could play an important role in methanogenesis. To systematically examine its physiological role, in this work, mmpX (locus tag MMP1554), the gene encoding Mmp10 in Methanococcus maripaludis, was deleted with a new genetic tool, resulting in the complete loss of the 5-C-(S)-methylarginine PTM of residue 275 in the McrA subunit. When the ΔmmpX mutant was complemented with the wild-type gene expressed by either a strong or a weak promoter, methylation was fully restored. Compared to the parental strain, maximal rates of methane formation by whole cells were reduced by 40 to 60% in the ΔmmpX mutant. The reduction in activity was fully reversed by the complement with the strong promoter. Site-directed mutagenesis of mmpX resulted in a differential loss of arginine methylation among the mutants in vivo, suggesting that activities of Mmp10 directly modulated methylation. R275 was present in a highly conservedmore » PXRR275(A/S)R(G/A) signature sequence in McrAs. The only other protein in M. maripaludis containing a similar sequence was not methylated, suggesting that Mmp10 is specific for McrA. In conclusion, Mmp10 modulates the methyl-Arg PTM on McrA in a highly specific manner, which has a profound impact on Mcr activity.« less

Authors:
 [1];  [1];  [1];  [1];  [1];  [2];  [1]
  1. Univ. of Georgia, Athens, GA (United States)
  2. Auburn Univ., AL (United States)
Publication Date:
Research Org.:
Univ. of Georgia, Athens, GA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES); National Science Foundation (NSF)
Contributing Org.:
University of Georgia iGEM Team, Korea Institute of Ocean Science and Technology
OSTI Identifier:
1595764
Grant/Contract Number:  
SC0018028; MCB-1410102
Resource Type:
Accepted Manuscript
Journal Name:
Journal of Bacteriology
Additional Journal Information:
Journal Volume: 202; Journal Issue: 3; Journal ID: ISSN 0021-9193
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; 03 NATURAL GAS; 09 BIOMASS FUELS; coenzyme M methylreductase; Methanococcus; post-translational modification; radical SAM enzyme

Citation Formats

Lyu, Zhe, Shao, Nana, Chou, Chau-Wen, Shi, Hao, Patel, Ricky, Duin, Evert C., and Whitman, William B. Posttranslational Methylation of Arginine in Methyl Coenzyme M Reductase Has a Profound Impact on both Methanogenesis and Growth of Methanococcus maripaludis. United States: N. p., 2019. Web. doi:10.1128/JB.00654-19.
Lyu, Zhe, Shao, Nana, Chou, Chau-Wen, Shi, Hao, Patel, Ricky, Duin, Evert C., & Whitman, William B. Posttranslational Methylation of Arginine in Methyl Coenzyme M Reductase Has a Profound Impact on both Methanogenesis and Growth of Methanococcus maripaludis. United States. https://doi.org/10.1128/JB.00654-19
Lyu, Zhe, Shao, Nana, Chou, Chau-Wen, Shi, Hao, Patel, Ricky, Duin, Evert C., and Whitman, William B. Mon . "Posttranslational Methylation of Arginine in Methyl Coenzyme M Reductase Has a Profound Impact on both Methanogenesis and Growth of Methanococcus maripaludis". United States. https://doi.org/10.1128/JB.00654-19. https://www.osti.gov/servlets/purl/1595764.
@article{osti_1595764,
title = {Posttranslational Methylation of Arginine in Methyl Coenzyme M Reductase Has a Profound Impact on both Methanogenesis and Growth of Methanococcus maripaludis},
author = {Lyu, Zhe and Shao, Nana and Chou, Chau-Wen and Shi, Hao and Patel, Ricky and Duin, Evert C. and Whitman, William B.},
abstractNote = {Catalyzing the key step for anaerobic production and/or oxidation of methane and likely other short-chain alkanes, methyl coenzyme M reductase (Mcr) and its homologs play a key role in the global carbon cycle. The McrA subunit possesses up to five conserved posttranslational modifications (PTMs) at its active site. It was previously suggested that methanogenesis marker protein 10 (Mmp10) could play an important role in methanogenesis. To systematically examine its physiological role, in this work, mmpX (locus tag MMP1554), the gene encoding Mmp10 in Methanococcus maripaludis, was deleted with a new genetic tool, resulting in the complete loss of the 5-C-(S)-methylarginine PTM of residue 275 in the McrA subunit. When the ΔmmpX mutant was complemented with the wild-type gene expressed by either a strong or a weak promoter, methylation was fully restored. Compared to the parental strain, maximal rates of methane formation by whole cells were reduced by 40 to 60% in the ΔmmpX mutant. The reduction in activity was fully reversed by the complement with the strong promoter. Site-directed mutagenesis of mmpX resulted in a differential loss of arginine methylation among the mutants in vivo, suggesting that activities of Mmp10 directly modulated methylation. R275 was present in a highly conserved PXRR275(A/S)R(G/A) signature sequence in McrAs. The only other protein in M. maripaludis containing a similar sequence was not methylated, suggesting that Mmp10 is specific for McrA. In conclusion, Mmp10 modulates the methyl-Arg PTM on McrA in a highly specific manner, which has a profound impact on Mcr activity.},
doi = {10.1128/JB.00654-19},
journal = {Journal of Bacteriology},
number = 3,
volume = 202,
place = {United States},
year = {Mon Nov 18 00:00:00 EST 2019},
month = {Mon Nov 18 00:00:00 EST 2019}
}

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