Reprint of: Inhibition of Escherichia coli RNAp by T7 Gp2 protein: Role of Negatively Charged Strip of Amino Acid Residues in Gp2

Carol Sheppard, Beatriz Cámara, Andrey Shadrin, Natalia Akulenko, Minhao Liu, Geoff Baldwin, Konstantin Severinov, Ernesto Cota, Steve Matthews, Siva R. Wigneshweraraj

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3 Citations (Scopus)

Abstract

Gp2, a 7 kDa protein encoded by T7 bacteriophage, is a potent inhibitor of Escherichia coli RNA polymerase (RNAp), the enzyme responsible for transcription of all bacterial genes and early viral genes. A prominent feature in the structure of Gp2 is a contiguous strip of seven negatively charged amino acid residues (negatively charged strip or NCS), located along one side of the molecule. The role of the NCS in Gp2 function is not known. Here, the in vivo and in vitro properties of altered forms of Gp2 with amino acid substitutions in the NCS are described. While mutations in the NCS do not compromise the folding or the ability of Gp2 to bind to the RNAp β′ subunit, disruption of the NCS significantly attenuates Gp2 function in vivo and its ability to inhibit RNAp in vitro. Efficient inhibition of the RNAp by Gp2 also involves the amino terminal region 1 domain of the RNAp promoter specificity subunit σ70, located in the vicinity of the primary Gp2 binding site in β′. The results are discussed in the context of hypothetical molecular mechanisms of RNAp inhibition by Gp2. © 2011 Elsevier Ltd. All rights reserved.
Original languageEnglish
Pages (from-to)832-841
Number of pages10
JournalJournal of Molecular Biology
DOIs
Publication statusPublished - 7 Oct 2011
Externally publishedYes

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    Sheppard, C., Cámara, B., Shadrin, A., Akulenko, N., Liu, M., Baldwin, G., Severinov, K., Cota, E., Matthews, S., & Wigneshweraraj, S. R. (2011). Reprint of: Inhibition of Escherichia coli RNAp by T7 Gp2 protein: Role of Negatively Charged Strip of Amino Acid Residues in Gp2. Journal of Molecular Biology, 832-841. https://doi.org/10.1016/j.jmb.2011.07.064