PMID: 9435223Jan 22, 1998Paper

Promoter specificity determinants of T7 RNA polymerase

Proceedings of the National Academy of Sciences of the United States of America
M RongR K Durbin

Abstract

The high specificity of T7 RNA polymerase (RNAP) for its promoter sequence is mediated, in part, by a specificity loop (residues 742-773) that projects into the DNA binding cleft (1). Previous work demonstrated a role for the amino acid residue at position 748 (N748) in this loop in discrimination of the base pairs (bp) at positions -10 and -11 (2). A comparison of the sequences of other phage RNAPs and their promoters suggested additional contacts that might be important in promoter recognition. We have found that changing the amino acid residue at position 758 in T7 RNAP results in an enzyme with altered specificity for the bp at position -8. The identification of two amino acid:base pair contacts (i.e., N748 with the bp at -10 and -11, and Q758 with the bp at -8) provides information concerning the disposition of the specificity loop relative to the upstream region of the promoter. The results suggest that substantial rearrangements of the loop (and/or the DNA) are likely to be required to allow these amino acids to interact with their cognate base pairs during promoter recognition.

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Citations

Feb 19, 2000·Current Opinion in Structural Biology·G M Cheetham, T A Steitz
Nov 8, 2002·Nature·Tahir H TahirovShigeyuki Yokoyama
Mar 13, 2013·Proceedings of the National Academy of Sciences of the United States of America·David L Shis, Matthew R Bennett
May 16, 2013·Proceedings of the National Academy of Sciences of the United States of America·Bryan C DickinsonDavid R Liu
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Aug 9, 2016·Plant Molecular Biology·Alexandra-Viola BohneThomas Börner
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Mar 21, 2009·PLoS Computational Biology·Oscar Westesson, Ian Holmes

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