       Document 0719
 DOCN  M9590719
 TI    Probing the details of the HIV-1 Rev-Rev-responsive element interaction:
       effects of modified nucleotides on protein affinity and conformational
       changes during complex formation.
 DT    9509
 AU    Renwick SB; Critchley AD; Adams CJ; Kelly SM; Price NC; Stockley PG;
       Department of Genetics, University of Leeds, U.K.
 SO    Biochem J. 1995 Jun 1;308 ( Pt 2):447-53. Unique Identifier : AIDSLINE
       MED/95289974
 AB    The solution structure of the human immunodeficiency virus type 1
       (HIV-1) Rev-responsive element (RRE) has been investigated by enzymic
       and chemical structural probing of a 71 nt RRE transcript. The minimum
       sequence information required to maintain recognition by the Rev protein
       has previously been mapped to a 29 nt stem-loop structure, known as
       minSLIIB. The key recognition target is a single-stranded RNA bubble at
       the base of the RNA stem. The fine details of RNA recognition have been
       probed using chemically synthesized minSLIIBs containing variant base or
       sugar residues at sites within the bubble. These have been analysed by
       gel retardation assays and their relative affinities for Rev protein
       determined. Complex formation between the wild-type minSLIIB RRE and Rev
       protein was also monitored using CD spectroscopy, which suggests a
       change in RNA conformation upon Rev binding. The spectral change is
       consistent with localized melting of RNA, leading to a decrease in the
       level of base stacking and/or a change in base tilting, during formation
       of the complex. Deoxynucleotide substitutions on just one side, the 5'
       side, of the bubble inhibit the conformational change detected by CD.
       The data are consistent with a dynamic interaction between Rev and its
       target site. The contact points between Rev and the RRE were probed
       directly using photo-cross-linking with either ribo-5-bromouridine- or
       ribo-4-thiouridine-substituted minSLIIBs. The data are consistent with
       protein-RNA contacts at the bottom of the bubble.
 DE    Base Sequence  Circular Dichroism  Gene Products, rev/*CHEMISTRY
       Hydrogen Bonding  HIV-1/*CHEMISTRY/GENETICS  Molecular Sequence Data
       Nucleic Acid Conformation  Photochemistry  Protein Binding  RNA-Binding
       Proteins/*CHEMISTRY  RNA, Viral/*CHEMISTRY  Structure-Activity
       Relationship  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).

