       Document 0450
 DOCN  M9440450
 TI    Viral long terminal repeat substrate binding characteristics of the
       human immunodeficiency virus type 1 integrase.
 DT    9404
 AU    Hazuda DJ; Wolfe AL; Hastings JC; Robbins HL; Graham PL; LaFemina RL;
       Emini EA; Merck Research Laboratories, West Point, Pennsylvania 19486.
 SO    J Biol Chem. 1994 Feb 11;269(6):3999-4004. Unique Identifier : AIDSLINE
       MED/94140811
 AB    A DNA binding assay was developed for the human immunodeficiency virus
       type 1 (HIV-1) integrase. The assay was capable of defining discrete
       complexes between the enzyme and the viral long terminal repeat (LTR)
       substrate. DNA binding reflected the sequence requirements previously
       demonstrated for the enzyme's 3'-end processing activity. Binding
       exhibited a nonlinear dependence on integrase concentration, suggesting
       that the enzyme functions as a multimer. The oligomeric state was
       investigated by UV-photo-cross-linking of integrase-LTR oligonucleotide
       complexes using DNA substrates substituted with 5-bromo-2'-deoxycytidine
       within the integrase recognition sequence. In the absence of divalent
       cation, integrase cross-linked to the LTR oligonucleotide as a single
       species whose mobility by SDS-polyacrylamide gel electrophoresis was
       consistent with the formation of tetramers. Using these techniques,
       analysis of the binding properties of integrase mutants demonstrated
       that the catalytic and sequence-specific DNA binding activities of the
       enzyme are distinct, involving residues within the conserved DD(35)E and
       zinc finger motifs, respectively.
 DE    Base Sequence  Cations, Divalent  DNA
       Nucleotidyltransferases/*METABOLISM  DNA-Binding Proteins/METABOLISM
       *HIV Long Terminal Repeat  HIV-1/*ENZYMOLOGY  Macromolecular Systems
       Molecular Sequence Data  Recombinant Proteins  Structure-Activity
       Relationship  Substrate Specificity  Zinc Fingers  JOURNAL ARTICLE

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

