PMID: 7020764Aug 27, 1981Paper

Spectroscopic studies of Congo Red binding to RNA polymerase

Biochimica Et Biophysica Acta
A Y WoodyR W Woody

Abstract

The azo dye Congo Red has a high affinity for nucleotide-binding enzymes. We have studied the binding of Congo Red to RNA polymerase by circular dichroism (CD) and difference absorption spectroscopy, steady-state kinetics, and nitrocellulose filter-binding. Induced CD shows that a large number of Congo Red molecules bind to the holoenzyme. CD also demonstrates that the core enzyme at low ionic strengths has a distinctive Congo Red binding site which is not present in the holoenzyme, nor in the core enzyme at higher ionic strengths or in the presence of poly(dT). CD studies indicate that Congo Red can readily displace double-stranded polynucleotides (T7 DNA or poly[d(A-T)] from RNA polymerase. Single-stranded DNA (poly(dT) and T7 DNA in open complexes) is not displaced from RNA polymerase except at high Congo Red concentrations. Both kinetics and nitrocellulose filter-binding measurements support this conclusion. Difference spectra indicate that the bound Congo Red molecules undergo stacking. We postulate that RNA polymerase binds Congo Red in a region with which a segment of DNA normally interacts, and that Congo Red is a potent inhibitor because the stacked dye has a polyanionic character.

References

Nov 21, 1977·Biochemical and Biophysical Research Communications·R A Edwards, R W Woody
Jan 1, 1972·Annual Review of Biochemistry·P H Von Hippel, J D McGhee
Jan 1, 1974·Annual Review of Biochemistry·M J Chamberlin
Oct 17, 1972·Biochemical and Biophysical Research Communications·E A Smuckler
Jun 1, 1974·The Biochemical Journal·R Eisenthal, A Cornish-Bowden
Dec 22, 1970·Biochemistry·D Berg, M Chamberlin
Apr 1, 1970·Proceedings of the National Academy of Sciences of the United States of America·A N Glazer
Aug 1, 1962·Journal of Molecular Biology·R B INMAN, R L BALDWIN
Apr 19, 1965·Biochimica Et Biophysica Acta·J S KRAKOW

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Citations

Sep 9, 2006·Brain Research Reviews·Petrea FridNatalija Popovic
Aug 1, 1992·Journal of Neurochemistry·B Caughey, R E Race
Jun 19, 2001·The Journal of Biological Chemistry·R KhuranaA L Fink
Jun 11, 2021·Journal of Computer-aided Molecular Design·Mariangela AgamennoneRoberto Paciotti

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