In vitro bypass of UV-induced lesions by Escherichia coli DNA polymerase I: specificity of nucleotide incorporation

Journal Article · Tue Mar 01 04:00:00 EST 1983 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.80.6.1541· OSTI ID:5724330

Rabkin, S D; Moore, P D; Strauss, B D

A variety of DNA polymerases, synthesizing in vitro on an UV-irradiated phiX174 DNA template, terminate synthesis one nucleotide before the 3' pyrimidines of putative dimers on the template. We have devised a system using Escherichia coli DNA polymerase I (Klenow fragment) that can synthesize past at least some of these dimers. The bypass is carried out in a multistep process--first, the incorporation of nucleotides opposite the pyrimidines in the dimer and, then, the addition of nucleotides complementary to the bases distal to the dimer. The insertion of a nucleotide opposite the first (3') pryimidine of a putative dimer in the presence of Mn/sup 2 +/ occurs in a concentration-dependent fashion with a 3- to 4-fold preference for purine nucleotides over pyrimidine nucleotides. In the presence of Mg/sup 2 +/, insertion is less frequent. Correlation of these results with in vivo mutation data suggests a role for the polymerase in determining the spectrum of base substitution mutagenesis in SOS induced cells.

• Research Organization: University of Chicago, IL
• DOE Contract Number: AC02-76EV02040
• OSTI ID: 5724330
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 80:6; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
ALKALINE EARTH METAL COMPOUNDS
AZINES
BACTERIA
BIOLOGICAL EFFECTS
BIOLOGICAL RADIATION EFFECTS
BIOSYNTHESIS
CATIONS
CHARGED PARTICLES
DIMERS
DNA
ELECTROMAGNETIC RADIATION
ENZYMES
ESCHERICHIA COLI
HETEROCYCLIC COMPOUNDS
IN VITRO
IONS
MAGNESIUM COMPOUNDS
MANGANESE COMPOUNDS
MICROORGANISMS
MUTAGENESIS
NUCLEIC ACIDS
NUCLEOTIDES
NUCLEOTIDYLTRANSFERASES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
PYRIMIDINES
RADIATION EFFECTS
RADIATIONS
SPECIFICITY
SYNTHESIS
TRANSFERASES
TRANSITION ELEMENT COMPOUNDS
ULTRAVIOLET RADIATION