Title

Footprinting, Circular Dichroism and UV Melting Studies on Neomycin B Binding to the Packaging Region of Human Immunodeficiency Virus Type-1 RNA

Author(s)/Creator(s)

Mark P. McPike, Syracuse University
Julie M. Sullivan, Syracuse University
Jerry Goodisman, Syracuse UniversityFollow
James C. Dabrowiak, Syracuse UniversityFollow

Document Type

Article

Date

5-9-2002

Keywords

article; binding affinity; binding site; circular dichroism; concentration response; controlled study; drug binding; drug specificity; Human immunodeficiency virus 1; measurement; melting point; molecular model; nonhuman; priority journal; protein footprinting; RNA structure; structure analysis; temperature measurement; ultraviolet spectrophotometry; virus strain; dimer; framycetin; nucleotide; pancreatic ribonuclease; virus RNA

Disciplines

Chemistry

Description/Abstract

We have studied the binding of neomycin to a 171mer RNA (Ψ-RNA) from the packaging region of the LAI strain of human immunodeficiency virus type-1, HIV-1 (LAI). The RNase I footprinting studies reveal that the primary binding site for the drug is in stem-loop 1, which contains the dimer initiation site of HIV-1. Loading this site with neomycin causes a structural change in the RNA, allowing nucleotides in the neighboring stem-loop 2 to participate in the drug site. Drug binding to secondary sites induces structural changes in other stem-loops of the RNA. Footprinting plots, showing cutting at a site as a function of drug concentration, were analyzed using a two-state model to obtain relative site-specific binding constants. Circular dichroism measurements show that neomycin binding to Ψ-RNA changes the intensity of the strong negative CD band at 208 nm, confirming that neomycin induces structural changes. Melting studies of the RNA showed melting transitions in the absence of drug at 28.2, 37.2, 47.4, 55.5 and 60.8°C. Only the first two were affected by drug binding, the reason for this being explained by our analysis.

Additional Information

Copyright 2002 Nucleic Acids Research. This article may be downloaded for personal use only. Any other use requires prior permission of the author and Nucleic Acids Research.

The article may be found at http://www.ncbi.nlm.nih.gov/pmc/articles/PMC117057/

Recommended Citation

McPike, Mark P.; Sullivan, Julie M.; Goodisman, Jerry; and Dabrowiak, James C., "Footprinting, Circular Dichroism and UV Melting Studies on Neomycin B Binding to the Packaging Region of Human Immunodeficiency Virus Type-1 RNA" (2002). Chemistry - All Scholarship. 18.
https://surface.syr.edu/che/18

Source

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Creative Commons License

This work is licensed under a Creative Commons Attribution 3.0 License.