PMID: 2125482Oct 23, 1990Paper

Circular dichroism studies of the HIV-1 Rev protein and its specific RNA binding site

Biochemistry
T J DalyA D Frankel

Abstract

The circular dichroism (CD) spectrum of the Rev protein from HIV-1 indicates that Rev contains about 50% alpha helix and 25% beta sheet at 5 degrees C in potassium phosphate buffer, pH 3, and 300 mM KF. The spectrum is independent of protein concentration over a 20-fold range. At neutral pH, Rev is relatively insoluble but can be brought into solution by binding to its specific RNA binding site, the Rev-responsive element (RRE), at a Rev:RNA ratio of about 3:1. Nonspecific binding to tRNA does not solubilize Rev. As judged by difference CD spectra, the conformation of Rev when bound to the RRE at neutral pH is similar to the conformation of unbound Rev at pH 3, although changes in the RNA may also contribute to the difference spectrum. Indeed, some difference is observed near 260 nm, consistent with a conformational change of the RRE upon Rev binding. Rev alone at pH 3 shows irreversible aggregation as the temperature is raised, while Rev bound to the RRE at neutral pH shows a reversible transition with a Tm of 68 degrees C.

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Citations

Jan 12, 2000·Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy·W ThumbM Auer
Feb 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·J KjemsP A Sharp
Apr 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·J Tao, A D Frankel
Jan 31, 2013·Nucleic Acids Research·Charlotte A HendersonAnastasia J Callaghan
Jan 19, 1999·Annual Review of Microbiology·V W Pollard, M H Malim
Jun 16, 2011·International Journal of Molecular Sciences·Vikas P Jain, Raymond S Tu
Sep 1, 2006·Current Opinion in HIV and AIDS·Dirk Daelemans, Christophe Pannecouque
Oct 15, 2008·Protein Expression and Purification·Marco MarenchinoMirko Hennig
Nov 23, 2011·Chembiochem : a European Journal of Chemical Biology·Yuan FuAnne M Baranger
Apr 14, 2011·Chembiochem : a European Journal of Chemical Biology·Peter SimanAshraf Brik
Jun 4, 1993·Cell·R TanA D Frankel
Mar 17, 2015·Biochemical and Biophysical Research Communications·Yuuki FujiwaraTomohiro Kabuta
May 1, 1997·Chemistry & Biology·T J Hope
Mar 30, 2001·The Journal of Biological Chemistry·M VilarE Perez-Paya
Jan 26, 2008·Protein Science : a Publication of the Protein Society·Stephen P EdgcombMirko Hennig
Aug 1, 1996·Journal of Virology·A Y Howe, D L Tyrrell
May 24, 1991·Science·B J CalnanA D Frankel

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