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Identification and Characterization of HEPN-MNT Type II TA System from Methanothermobacter thermautotrophicus ΔH

  • Microbial Genetics, Genomics and Molecular Biology
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Abstract

Toxin-antitoxin (TA) systems are widespread in bacteria and archaea plasmids and genomes to regulate DNA replication, gene transcription, or protein translation. Higher eukaryotic and prokaryotic nucleotide-binding (HEPN) and minimal nucleotidyltransferase (MNT) domains are prevalent in prokaryotic genomes and constitute TA pairs. However, three gene pairs (MTH304/305, 408/409, and 463/464) of Methanothermobacter thermautotropicus ΔH HEPN-MNT family have not been studied as TA systems. Among these candidates, our study characterizes the MTH463/MTH464 TA system. MTH463 expression inhibited Escherichia coli growth, whereas MTH464 did not and blocked MTH463 instead. Using site-directed MTH463 mutagenesis, we determined that amino acids R99G, H104A, and Y106A from the R[ɸX]4-6H motif are involved with MTH463 cell toxicity. Furthermore, we established that purified MTH463 could degrade MS2 phage RNA, whereas purified MTH464 neutralized MTH463 activity in vitro. Our results indicate that the endonuclease toxin MTH463 (encoding a HEPN domain) and its cognate antitoxin MTH464 (encoding the MNT domain) may act as a type II TA system in M. thermautotropicus ΔH. This study provides initial and essential information studying TA system functions, primarily archaea HEPN-MNT family.

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The datasets generated or analyzed during this study are available from the corresponding author on reasonable request.

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Acknowledgements

This research was carried out with the support of “Cooperative Research Program for Agriculture Science and Technology Development (Project No. PJ01595802)” Rural Development Administration, Republic of Korea, and of the Korea Research Institute of Bioscience and Biotechnology (KRIBB) Research Initiative Program (KGM5362322).

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  1. Wonho Choi

    Present address: 4D Convergence Technology Institute (National Key Technology Institute in University), Korea National University of Transportation, Jungpyeong, 27909, Republic of Korea

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  1. Bio-Evaluation Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju, 28116, Republic of Korea

    Wonho Choi, Anoth Maharjan, Ji-Young Park & Jung-Ho Park

  2. BIORCHESTRA Co., LTD., Daejeon, 34013, Republic of Korea

    Hae Gang Im

  3. Department of Food Science and Technology, Chungnam National University, Daejeon, 34134, Republic of Korea

    Jong-Tae Park

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Choi, W., Maharjan, A., Im, H.G. et al. Identification and Characterization of HEPN-MNT Type II TA System from Methanothermobacter thermautotrophicus ΔH. J Microbiol. 61, 411–421 (2023). https://doi.org/10.1007/s12275-023-00041-9

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