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Site-specifically modified oligodeoxyribonucleotides as templates for Escherichia coli DNA polymerase I

Journal Article · · Proc. Natl. Acad. Sci. U.S.A.; (United States)
;

Oligodeoxyribonucleotides with site-specific modifications have been used as substrates for Escherichia coli DNA polymerase I holoenzyme and Klenow fragment. Modifications included the bulky guanine-8-aminofluorene adduct and a guanine oxidation product resembling the product of photosensitized DNA oxidation. By a combination of primers and nick-mers, conditions of single-strand-directed DNA synthesis and nick-translation could be created. The results show that the polymerase can bypass both types of lesions. Bypass occurs on a single-stranded template but is facilitated on a nicked, double-stranded template. Only purines, with guanine more favored than adenine, are incorporated across both lesions. The results indicate that site-specifically modified oligonucleotides can be sensitive probes for the action of polymerases on damaged templates. They also suggest a function for polymerase I, in its nick-translation capacity, during DNA repair and mutagenesis.

Research Organization:
Memorial Sloan-Kettering Cancer Center, Rye, NY
OSTI ID:
6291080
Journal Information:
Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 82:8; ISSN PNASA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AROMATICS
AUTORADIOGRAPHY
AZAARENES
BACTERIA
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMISTRY
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
BIOSYNTHESIS
CHEMISTRY
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
DNA POLYMERASES
DNA REPAIR
DNA REPLICATION
ENZYMES
ESCHERICHIA COLI
HETEROCYCLIC COMPOUNDS
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
LIQUID COLUMN CHROMATOGRAPHY
MICROORGANISMS
NUCLEI
NUCLEIC ACID REPLICATION
NUCLEOTIDES
NUCLEOTIDYLTRANSFERASES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
PURINES
RADIOISOTOPES
RECOVERY
REPAIR
SEPARATION PROCESSES
SYNTHESIS
TRACER TECHNIQUES
TRANSFERASES