Site-specific hydrolysis and alcoholysis of human immunodeficiency virus DNA termini mediated by the viral integrase protein.
Before integration of the human immunodeficiency virus (HIV) DNA, two nucleotides are removed from the 3' ends of the viral DNA by the integrase (IN) protein. We studied the chemistry of this reaction, and found that IN mediates site-specific hydrolysis of a phosphodiester bond, resulting in release of a dinucleotide. A class of alcohols (including glycerol, 1,2-propanediol, but not 1,3-propanediol) can also act as nucleophile in this reaction, and likewise the alcoholic amino acids L-serine and L-threonine can be covalently linked to the dinucleotide. No evidence was found for a covalent linkage between the IN protein and this dinucleotide, suggesting that IN directs a single nucleophilic attack of water at the specific phosphodiester bond.
Vink
C
C
Division of Molecular Biology, The Netherlands Cancer Institute, Amsterdam.
Yeheskiely
E
E
van der Marel
G A
GA
van Boom
J H
JH
Plasterk
R H
RH
D001358:Avian Sarcoma Viruses / Q000378:metabolism; D001483:Base Sequence; D004254:DNA Nucleotidyltransferases / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D005990:Glycerol / Q000378:metabolism; D015497:HIV-1 / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D015498:HIV-2 / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D019426:Integrases; D008969:Molecular Sequence Data; D009711:Nucleotides / Q000378:metabolism; D019946:Propylene Glycol; D011409:Propylene Glycols / Q000378:metabolism; D012680:Sensitivity and Specificity; D012694:Serine / Q000378:metabolism; D013912:Threonine / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism*
D004279:DNA, Viral; D009711:Nucleotides; D011409:Propylene Glycols; D014764:Viral Proteins; D013912:Threonine; D012694:Serine; D019946:Propylene Glycol; D004254:DNA Nucleotidyltransferases; D019426:Integrases; D005990:Glycerol