Document 0504
 DOCN  M9590504
 TI    Intermolecular binding sites of human immunodeficiency virus type 1 Rev
       protein determined by protein footprinting.
 DT    9509
 AU    Jensen TH; Leffers H; Kjems J; Department of Molecular Biology,
       University of Aarhus, Denmark.
 SO    J Biol Chem. 1995 Jun 9;270(23):13777-84. Unique Identifier : AIDSLINE
       MED/95293973
 AB    Human immunodeficiency virus encodes the regulatory protein Rev, which
       is required for expression of viral structural proteins. It binds to an
       RNA element (RRE) in the viral transcript and up-regulates the
       cytoplasmic appearance of unspliced and singly spliced viral mRNA. We
       have studied the structure of Rev alone and complexed with the RRE and
       two monoclonal antibodies, using a protein footprinting approach. The
       method involves radioactive labeling at the C-terminal end of Rev fusion
       protein followed by limited proteolysis under native conditions, using
       10 different proteinases. Rev protein was mainly cleaved within the
       basic domain and in the C-terminal part. The periodicity of the
       proteolytic cleavages within the basic domain strongly suggests that it
       forms an alpha-helical structure with one side facing the solvent. In
       the presence of RRE, these cleavages became significantly reduced. In
       addition, strong protection was observed at position 66 outside the
       basic domain. As a control for the specificity of the footprinting
       reaction, we confirmed the position of the epitopes for two monoclonal
       antibodies. This protein footprinting methodology is generally
       applicable to other proteins for which terminal modifications are
       acceptable, and provides a useful tool for mapping structure, substrate
       binding, and conformational changes.
 DE    Base Sequence  Binding Sites  Epitope Mapping  Gene Products,
       rev/CHEMISTRY/IMMUNOLOGY/*METABOLISM  *HIV-1  Molecular Sequence Data
       Recombinant Fusion Proteins/METABOLISM  RNA/METABOLISM  Support,
       Non-U.S. Gov't  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).