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M94B0798.TXT
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1994-11-11
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Document 0798
DOCN M94B0798
TI Replication inhibition and miscoding properties of a DNA template
containing N-(deoxyguanosin-8-yl)-1-aminopyrene (Meeting abstract).
DT 9412
AU Vyas RR; Basu AK; Dept. of Chemistry, Univ. of Connecticut, Storrs, CT
06269
SO Proc Annu Meet Am Assoc Cancer Res; 35:A854 1994. Unique Identifier :
AIDSLINE ICDB/94602541
AB A major DNA adduct formed by the environmental pollutant 1-nitropyrene
is N-(deoxyguanosin-8-yl)-1-aminopyrene (dG(AP)). We have synthesized a
DNA fragment that contained this adduct at a specific site. A primer was
annealed to this template and in vitro DNA synthesis by a variety of
polymerases was studied. Primer extension catalyzed by HIV reverse
transcriptase, a modified T7 DNA polymerase (Sequenase), human DNA
polymerase alpha, or DNA polymerase beta was inhibited almost
quantitatively at the base 3' to the adduct site even when high
concentration of the polymerase and/or dNTPs were employed. When
3'----5' exonuclease-free Klenow fragment of the DNA polymerase I was
used, efficient nucleotide incorporation opposite the adduct was
observed. However, dGTP and dATP were preferentially incorporated
opposite dG(AP). In the presence of Mg2+, extension beyond the adduct
site did not occur. In the presence of Mn2+, on the other hand,
significant proportion of the primer was extended to a full-length
product. This suggests that dG(AP) can induce both genotoxic and
mutagenic effects in vivo.
DE Comparative Study DNA Polymerase I/METABOLISM DNA Polymerase
II/METABOLISM DNA Polymerases/*METABOLISM DNA Primers *DNA
Replication/DRUG EFFECTS Deoxyguanosine/*ANALOGS &
DERIVATIVES/METABOLISM Environmental Pollutants/*TOXICITY
Exodeoxyribonucleases/METABOLISM HIV/ENZYMOLOGY Human
Pyrenes/*ANALYSIS/METABOLISM/*TOXICITY Reverse
Transcriptase/*METABOLISM Templates MEETING ABSTRACT
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).