PMID: 6402775Feb 1, 1983Paper

Mammalian mutator mutant with an aphidicolin-resistant DNA polymerase alpha

Proceedings of the National Academy of Sciences of the United States of America
P K LiuL A Loeb

Abstract

The Chinese hamster V79 cell mutant aphr-4-2, selected for its resistance to aphidicolin, a specific inhibitor of DNA polymerase alpha (DNA nucleotidyltransferase, EC 2.7.7.7), is characterized by slow growth, UV sensitivity, and hypersensitivity to UV-induced mutation. DNA polymerase alpha has been purified from mitochondria-free crude extracts of the mutant and its parental wild-type cells by sequential column chromatography on DEAE-cellulose and phosphocellulose. The major DNA polymerase activity from both cell lines was found to have characteristics of the alpha-type polymerase: sensitivity to 0.2 M KCl, resistance to heat denaturation (45 degrees C for 15 min), an apparent Km of 5 microM for dATP, and an ability to copy poly(dT)X(rA)10 but not poly(rA)X(dT)12. The crude extracts and purified DNA polymerase alpha from the mutant cells are not inhibited by aphidicolin (greater than 0.6 microM). The apparent Km for dCTP with DNA polymerase alpha is 1.0 +/- 0.4 microM (mean +/- SD) for the mutant enzyme. The polymerase from the parental cells, similarly purified, is sensitive to aphidicolin and has an apparent Km for dCTP of 10 +/- 4 microM. The spontaneous mutation rate (per cell per division), determined by fluctuation analy...Continue Reading

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Citations

Jun 1, 1987·Molecular & General Genetics : MGG·S AizawaG M Martin
Sep 1, 1990·Somatic Cell and Molecular Genetics·M ItoD Ayusawa
Jan 1, 1991·Somatic Cell and Molecular Genetics·P K Liu, T H Norwood
May 1, 1985·Somatic Cell and Molecular Genetics·S AizawaG M Martin
Jul 1, 1985·Somatic Cell and Molecular Genetics·K FukuchiY Okada
May 1, 1984·Somatic Cell and Molecular Genetics·P K LiuJ E Trosko
Oct 1, 1983·Mutation Research·I C LiE H Chu
Jan 1, 1990·Pharmacology & Therapeutics·R V Talanian, G E Wright
Jun 1, 1985·Molecular and Biochemical Parasitology·J Inselburg
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Oct 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·G Seal, M A Sirover
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Sep 1, 1990·Mutation Research·F W Perrino, L A Loeb

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