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Early antitermination in the atypical coliphage mEp021 mediated by the Gp17 protein

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Abstract

The coliphage mEp021 belongs to a phage group with a unique immunity repressor, and its life cycle requires the host factor Nus. mEp021 has been classified as non-lambdoid based on its specific characteristics. The mEp021 genome carries a gene encoding an Nλ-like antiterminator protein, termed Gp17, and three nut sites (nutL, nutR1, and nutR2). Analysis of plasmid constructs containing these nut sites, a transcription terminator, and a GFP reporter gene showed high levels of fluorescence when Gp17 was expressed, but not in its absence. Like lambdoid N proteins, Gp17 has an arginine-rich motif (ARM), and mutations in its arginine codons inhibit its function. In infection assays using the mutant phage mEp021ΔGp17::Kan (where gp17 has been deleted), gene transcripts located downstream of transcription terminators were obtained only when Gp17 was expressed. In contrast to phage lambda, mEp021 virus particle production was partially restored (>1/3 relative to wild type) when nus mutants (nusA1, nusB5, nusC60, and nusE71) were infected with mEp021 and Gp17 was overexpressed. Our results suggest that RNA polymerase reads through the third nut site (nutR2), which is more than 7.9 kbp downstream of nutR1.

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Data availability

The datasets generated and analyzed during the current study are available from the corresponding author upon reasonable request.

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Acknowledgments

We appreciate the valuable, sharp, careful comments and observations by the anonymous reviewers. Also, we thank Dr. Jaime Ortega and Dr. Norma Oviedo for their invaluable comments throughout this study. We are grateful to M.Sc. Ma. Guadalupe Aguilar-Gonzalez and Dr. Dulce Maria del Carmen Delgadillo Alvarez from LaNse-Cinvestav for technical assistance in DNA sequencing. We also thank M.Sc. Jose Bueno-Martinez and M.Sc. Carlos Osorio for helpful technical assistance, and M.Sc. Jairo Hurtado-Cortes and Dr. Victor Flores for bioinformatic assistance. During this work, G.V.T. and E.P.B.T. were granted doctoral fellowships from CONACyT, Mexico (No. 627423) and (No. 535549), respectively.

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CINVESTAV supported this work.

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Authors and Affiliations

  1. Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, Mexico

    Guadalupe Valencia-Toxqui, Elissa P. Ballinas-Turrén, Rosa Ma. Bermúdez-Cruz, Eva Martínez-Peñafiel, Gabriel Guarneros & Luis Kameyama

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  1. Guadalupe Valencia-Toxqui
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  6. Luis Kameyama
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Contributions

GV-T did experimental and analysis work, wrote the first manuscript draft, and conceptualized the work. Supporting experimental work was performed by EPB-T. EM-P did genome sequence assembly. Manuscript editing, corrections, suggestions, and comments were made by RMB-C. Supervision, suggestions, comments, and advice were given by GG. LK contributed to conceptualization, management, direction, and manuscript writing. All authors have read and approved the final manuscript.

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Correspondence to Luis Kameyama.

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Valencia-Toxqui, G., Ballinas-Turrén, E.P., Bermúdez-Cruz, R.M. et al. Early antitermination in the atypical coliphage mEp021 mediated by the Gp17 protein. Arch Virol 168, 92 (2023). https://doi.org/10.1007/s00705-023-05721-w

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