Sites of termination of in vitro DNA synthesis on ultraviolet- and N-acetylaminofluorene-treated phi X174 templates by prokaryotic and eukaryotic DNA polymerases
In vitro DNA synthesis on a phi X174 template primed with a restriction fragment and catalyzed by the Escherichia coli DNA polymerase I large (Klenow) fragment (pol I) terminates at the nucleotide preceding a site that has been altered by ultraviolet irradiation or treatment with N-acetylaminofluorene. Termination on ultraviolet-irradiated templates is similar when synthesis is catalyzed by E. coli DNA polymerase III holoenzyme (pol III), phage T4DNA polymerase a polymerase ..cap alpha.. from human lymphoma cells, or avian myeloblastosis virus reverse transcriptase. 3' ..-->.. 5' exonuclease activity cannot be detected in the reverse transcriptase and DNA polymerase ..cap alpha.. preparations. On N-acetylaminofluorene templates, pol I, pol III, and T4 polymerase reactions terminate immediately preceding the lesion, whereas reverse transcriptase-catalyzed reactions and, at some positions in the sequence, polymerase ..cap alpha..-catalyzed reactions terminate at the site of the lesion. Substitution of Mn/sup 2 +/ for Mg/sup 2 +/ changes the pattern of pol I-catalyzed termination sites. The data sugest that termination is a complicated process that does not depend exclusively on the 3' ..-->.. 5' exonuclease activity associated with many polymerases.
- Research Organization:
- Univ. of Chicago, IL
- OSTI ID:
- 6865161
- Journal Information:
- Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 78:1; ISSN PNASA
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
AROMATICS
BACTERIA
BIOASSAY
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL EFFECTS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
CONDENSED AROMATICS
DATA
DNA
DNA REPLICATION
ELECTROMAGNETIC RADIATION
ENZYME ACTIVITY
ENZYMES
ESCHERICHIA COLI
ESTERASES
GENETIC EFFECTS
HYDROLASES
IN VITRO
INFORMATION
KINETICS
MICROORGANISMS
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NUCLEIC ACID REPLICATION
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PHOSPHODIESTERASES
RADIATIONS
REACTION KINETICS
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REPAIR
TRANSCRIPTION
ULTRAVIOLET RADIATION