       Document 0166
 DOCN  M9630166
 TI    High-affinity interaction of human immunodeficiency virus type-1 reverse
       transcriptase with partially complementary primers.
 DT    9603
 AU    Zakharova OD; Tarrago-Litvak L; Maksakova G; Andreola ML; Dufour E;
       Litvak S; Nevinsky GA; Novosibirsk Institute of Bioorganic Chemistry,
       Siberian Division; of the Russian Academy of Sciences, Russia.
 SO    Eur J Biochem. 1995 Nov 1;233(3):856-63. Unique Identifier : AIDSLINE
       MED/96085150
 AB    The comparison of Km and Vmax values for various primers in the reaction
       of polymerization catalyzed by the human immunodeficiency virus type-1
       (HIV-1) reverse transcriptase was carried out. The primers were: (a)
       complementary to the template, (b) partially complementary with
       mismatched nucleotides at different positions from the 3' end or (c)
       non-complementary. Non-complementary primers were not elongated by HIV-1
       reverse transcriptase. However, if they contained only one residue
       complementary to the template or an abasic unit at the 3' end, they
       could serve as primers. The most effective discrimination between
       matched and mismatched primers, due to a decrease in the affinity and
       Vmax, was found in the case of oligonucleotides containing
       non-complementary bases at the second or third position from the 3' end
       of the primer. The efficiency of discrimination by HIV-1 reverse
       transcriptase between matched and mismatched base-paired primers was
       about 1-1.5 orders of magnitude lower than that of procaryotic,
       eucaryotic and archaebacterial DNA polymerases and avian myeloblastosis
       virus reverse transcriptase. Oligonucleotides such as (dT)4(dCdG)k(dT)4
       showed higher affinity for the enzyme than (dT)4 or (dT)8 primers. These
       data suggest that HIV-1 reverse transcriptase, in contrast to
       procaryotic, eucaryotic and archaebacterial DNA polymerases, forms
       additional contacts with the 5'-end region of the non-complementary
       primer. In addition, using tRNA(3Lys), the natural primer of HIV-1, it
       was shown that the p66 subunit of reverse transcriptase can be
       crosslinked, in the presence of a platinum derivative, to the 5' end of
       tRNA. Thus, besides the normal binding site for the 3' end of tRNA,
       which is crucial for the initiation of cDNA synthesis, the 5' end of the
       tRNA also interacts with a specific site on the enzyme.
 DE    DNA, Complementary  Human  HIV-1/*ENZYMOLOGY  Kinetics
       Oligonucleotides/GENETICS/*METABOLISM  RNA-Directed DNA
       Polymerase/GENETICS/*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).

