Selection by phage display of peptides targeting the HIV-1 TAR element

RNA Biol. 2005 Jan;2(1):28-33. doi: 10.4161/rna.2.1.1681. Epub 2005 Jan 25.

Abstract

As transcription regulatory element, the HIV-1 TAR RNA element is a promising target to inhibit viral replication; indeed, ligands of TAR RNA could prevent the transcription trans-activation process. Phage display in vitro selection was undertaken to select peptidic ligands of TAR RNA. In preliminary experiments, the selection was performed in a magnesium rich buffer (3 mM), but only phages targeted to plastic wells or streptavidin emerged; in addition, a "super-infectious" phage present in the New England Biolabs library (SVSVGMKPSPRP) selected by others with different targets was cloned, due to a high amplification potential. In contrast, the absence of magnesium or an increasing magnesium concentration (0 to 0.5 mM) led to phage selection with 57 amino acid peptides. K(D)s of 420-550 nM were measured by filter binding assays; a significant specificity was obtained when TAR target was compared with unrelated RNA targets. Surprisingly, the binding of selected peptides does not depend on the magnesium concentration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Buffers
  • HIV Long Terminal Repeat*
  • HIV-1 / genetics*
  • HIV-1 / physiology
  • Magnesium
  • Nucleic Acid Conformation
  • Peptide Library*
  • RNA-Binding Proteins / metabolism
  • Virus Replication*

Substances

  • Buffers
  • Peptide Library
  • RNA-Binding Proteins
  • Magnesium