PMID: 8450553Mar 5, 1993Paper

High affinity binding of TAR RNA by the human immunodeficiency virus type-1 tat protein requires base-pairs in the RNA stem and amino acid residues flanking the basic region

Journal of Molecular Biology
M J ChurcherJ Karn

Abstract

The binding site for tat protein on TAR RNA has been defined in quantitative terms using an extensive series of mutations. The relative dissociation constants for the mutant TAR RNAs were measured using a dual-label competition filter binding assay in which 35S-labelled wild-type TAR RNA (K1) was competed against 3H-labelled mutant TAR RNA (K2). The error in the self-competition experiment was usually less than 10% (e.g. K2/K1 = 1.07 +/- 0.05, n = 19) and the experimental data accurately matched theoretical curves calculated with fitted dissociation constants. Mutations in U23, a critical residue in the U-rich "bulge" sequence, or in either of the two base-pairs immediately above the "bulge", G26.C39 and A27.U38 reduced that affinity by 8- to 20-fold. Significant contributions to tat binding affinity were also made by the base-pairs located immediately below the bulge. For example, mutation of A22.U40 to U.A reduced tat affinity 5-fold, and mutation of G21.C41 to C.G reduced tat affinity 4-fold. The binding of a series of peptides spanning the basic "arginine-rich" sequence of tat was examined using both filter-binding and gel mobility shift assays. Each of the peptides showed significantly reduced affinities for wild-type TAR ...Continue Reading

Citations

Nov 26, 2010·Journal of the American Chemical Society·Wei HuangGary P Drobny
Jan 1, 1996·Nature Structural Biology·W I Sundquist
Nov 14, 1997·Nature Structural Biology·I HuqT M Rana
Jul 9, 2009·Proceedings of the National Academy of Sciences of the United States of America·Amy DavidsonGabriele Varani
Jul 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·M A GraebleJ Karn
May 23, 1995·Proceedings of the National Academy of Sciences of the United States of America·L Chen, A D Frankel
Jun 6, 1995·Proceedings of the National Academy of Sciences of the United States of America·R Tan, A D Frankel
Jun 20, 1995·Proceedings of the National Academy of Sciences of the United States of America·M Zacharias, P J Hagerman
Oct 1, 1996·Proceedings of the National Academy of Sciences of the United States of America·R K MishraJ J Toulmé
Mar 19, 1996·Proceedings of the National Academy of Sciences of the United States of America·N J KeenJ Karn
Mar 3, 1999·Journal of Biomolecular Structure & Dynamics·M J SeewaldH Sticht
Mar 26, 2010·AIDS Research and Human Retroviruses·Zhiming FangYiming Shao
Nov 11, 1993·Nucleic Acids Research·B Berkhout, B Klaver
Aug 25, 1994·Nucleic Acids Research·B BakerS M Kingsman
May 11, 1995·Nucleic Acids Research·R S DevonA J Brookes
Oct 1, 1996·Nucleic Acids Research·P R BohjanenM A Garcia-Blanco
Jun 5, 1998·Nucleic Acids Research·A GarbesiT Klimkait
Jun 6, 1998·Nucleic Acids Research·A S BrodskyJ R Williamson
Jan 11, 2000·Nucleic Acids Research·R NifosìP A Kollman
Jun 18, 2005·Nucleic Acids Research·Greg L OlsenGary P Drobny
Feb 23, 2012·Cold Spring Harbor Perspectives in Medicine·Jonathan Karn, C Martin Stoltzfus
Jun 29, 1999·Cold Spring Harbor Symposia on Quantitative Biology·M E GarberK A Jones
Sep 15, 1996·European Journal of Biochemistry·S M Kingsman, A J Kingsman

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