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Genetic analysis of bacteriophage λN-dependent antitermination suggests a possible role for the RNA polymerase α subunit in facilitating specific functions of NusA and NusE

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Abstract

A role for the Escherichia coli RNA polymerase α subunit in transcription antitermination dependent on bacteriophage λ N protein has been previously inferred from the isolation of rpoA mutants that alter the efficiency of this process. This report describes studies on the efficiency of N-dependent transcription antitermination in a strain containing the rpoA341 mutation, which interferes with this process. The effect of mutations in genes coding for different Nus factors and/or plasmids overexpressing nus genes on bacteriophage λ development in an E. coli rpoA341 host was examined. In addition, the effect of overproduction of the N protein in these genetic backgrounds was assessed. Analogous bacterial strains were employed to measure the efficiency of the antitermination process using the lacZ reporter gene under control of the λ p R promoter, and containing the phage nutR region and the t R1 terminator between the promoter and lacZ. The experimental results suggest interactions between components of the N-antitermination complex, which have been established biochemically, as well as additional functional relationships within the complex. Furthermore, the results indicate that amino acid substitution in the α subunit C-terminal domain encoded by the rpoA341 mutation may specifically disrupt the function of the NusA and NusE proteins. During this analysis, it was also found that the E. coli nusA1 mutant exhibits a conditional lethal phenotype.

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Acknowledgements

This work was supported by the Polish State Committee for Scientific Research (grant no. 3P04A04924 to G.W.) and the Wellcome Trust (grant 050794 to M.S.T.). G.W. also acknowledges financial support from the Foundation for Polish Science (subsidy 142000).

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Authors and Affiliations

  1. Department of Molecular Biology, University of Gdańsk, Kładki 24, 80–822, Gdańsk, Poland

    Agnieszka Szalewska-Pałasz, Barbara Strzelczyk, Anna Herman-Antosiewicz & Grzegorz Węgrzyn

  2. Institute of Oceanology, Polish Academy of Sciences, Św. Wojciecha 5, 81–347 , Gdynia, Poland

    Grzegorz Węgrzyn

  3. Division of Genomic Medicine, University of Sheffield Medical School, Beech Hill Road, Sheffield , S10 2RX, UK

    Mark S. Thomas

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  1. Agnieszka Szalewska-Pałasz
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Correspondence to Mark S. Thomas.

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Szalewska-Pałasz, A., Strzelczyk, B., Herman-Antosiewicz, A. et al. Genetic analysis of bacteriophage λN-dependent antitermination suggests a possible role for the RNA polymerase α subunit in facilitating specific functions of NusA and NusE. Arch Microbiol 180, 161–168 (2003). https://doi.org/10.1007/s00203-003-0571-2

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