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Effect of pH on the base-mispairing properties of 5-bromouracil during DNA synthesis

Journal Article · · Biochemistry; (United States)
DOI:https://doi.org/10.1021/bi00405a052· OSTI ID:5381933
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The authors have utilized an electrophoretic assay of misincorporation and autoradiography to investigate the possibility that ionization of 5-bromouracil (BU) may play a role in its mispairing during DNA synthesis in vitro. They examined the effects of increasing pH on the relative rates of formation of BU x G and T x G mispairs during chain elongation catalyzed by various DNA polymerases. For the Klenow fragment of Escherichia coli DNA polymerase I, increasing pH facilitated BU x G mispair formation (relative to T x G mispairing) when BU was present in the template strand. This effect showed a strong dependence on sequence context. Increasing pH had little effect on the relative rate of misincorporation of BrdUMP versus dTMP (at template G) by the Klenow polymerase. Misincorporation opposite template BU residues catalyzed by Maloney murine leukemia virus DNA polymerase and DNA polymerase ..beta.. (Novikoff hepatoma) also increased with pH, but for these two enzymes, there was no apparent dependence on sequence context. With T4 DNA polymerase and E. coli DNA polymerase III holoenzyme, a similar occurrence of BU x G and T x G mispairing during polymerization was observed, whether BU was present in the template or in the incoming nucleotide, and there was little effect of pH. The results reported here are consistent with a mispairing mechanism for template BU wherein the anionic form of the base mispairs with G.

Research Organization:
Baylor College of Medicine, Houston, TX (USA)
OSTI ID:
5381933
Journal Information:
Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 27:5; ISSN BICHA
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANTIMETABOLITES
ATP
AUTORADIOGRAPHY
AZINES
BACTERIOPHAGES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMISTRY
BIOLOGICAL EFFECTS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
BROMOURACILS
CHEMISTRY
DAYS LIVING RADIOISOTOPES
DNA POLYMERASES
DNA REPAIR
DNA REPLICATION
DRUGS
ELECTROPHORESIS
ENZYMES
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
ISOTOPES
LIGHT NUCLEI
MICROORGANISMS
NUCLEI
NUCLEIC ACID REPLICATION
NUCLEOTIDES
NUCLEOTIDYLTRANSFERASES
ODD-ODD NUCLEI
ORGANIC BROMINE COMPOUNDS
ORGANIC COMPOUNDS
ORGANIC HALOGEN COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PARASITES
PH VALUE
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
PYRIMIDINES
RADIOISOTOPES
RECOVERY
REPAIR
TRANSFERASES
URACILS
VIRUSES