       Document 0425
 DOCN  M9590425
 TI    Isolation of high-affinity RNA ligands to HIV-1 integrase from a random
       pool.
 DT    9509
 AU    Allen P; Worland S; Gold L; Department of Molecular, Cellular and
       Developmental Biology,; University of Colorado at Boulder 80309-0347,
       USA.
 SO    Virology. 1995 Jun 1;209(2):327-36. Unique Identifier : AIDSLINE
       MED/95297133
 AB    We were able to isolate high-affinity RNAs from a random pool that binds
       to integrase protein from the human immunodeficiency virus-type 1 using
       the procedure now known as SELEX. Generally, the RNAs fell into three
       different classes in binding buffer containing 250 mM NaCl: group I
       class of molecules binds integrase with a dissociation constant (Kd) on
       the order of 10 nM, group II molecules had a Kd of about 80 nM, and
       group III about 800 nM. The RNA with the highest affinity from the group
       I class of molecules, designated P5, was characterized using computer
       modeling, chemical and enzymatic probing, and deletion analysis. Our
       secondary structure model for this RNA suggests interactions between
       looped-out fixed nucleotides and nucleotides from the randomized region;
       a GNRA tetraloop is also in the structure. We showed that our integrase
       was able to process a U5 mimic in vitro. P5 competes effectively for
       binding with the double-stranded DNA mimic of U5 at 180 mM NaCl
       concentration.
 DE    Base Sequence  Cloning, Molecular  Comparative Study  Computer
       Simulation  DNA Nucleotidyltransferases/GENETICS/ISOLATION & PURIF/
       *METABOLISM  DNA Primers  Genes, Structural, Viral
       HIV-1/*ENZYMOLOGY/GENETICS  Kinetics  Ligands  Models, Molecular
       Molecular Sequence Data  Nucleic Acid Conformation  Polymerase Chain
       Reaction  Protein Binding  Recombinant Proteins/GENETICS/ISOLATION &
       PURIF/METABOLISM  RNA, Viral/CHEMISTRY/ISOLATION & PURIF/*METABOLISM
       Substrate Specificity  Support, Non-U.S. Gov't  Support, U.S. Gov't,
       P.H.S.  Virus Integration  JOURNAL ARTICLE

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

