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Mammalian DNA polymerase
Volume 270,
Number 27,
Issue of July 07, pp. 16402-16408, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
by DNA-alkylating
Agents in Chinese Hamster Ovary Cells
(-pol), a DNA repair
polymerase, is known to be constitutively expressed in cultured cells,
but treatment of cells with the DNA-alkylating agents MNNG or methyl
methanesulfonate has been shown to up-regulate -pol mRNA level. To
further characterize this response, we prepared a panel of monoclonal
antibodies and used one of them to quantify -pol in whole cell
extracts by immunoblotting. We found that treatment of Chinese hamster
ovary cells with either DNA-alkylating agent up-regulated the -pol
protein level 5-10-fold. This induction appeared to be secondary
to DNA alkylation, as induction was not observed with a genetically
altered cell line overexpressing the DNA repair enzyme O
-methylguanine-methyltransferase. We also found
that 12-O-tetradecanoylphorbol-13-acetate (TPA) treatment of
wild type Chinese hamster ovary cells increased expression of -pol
protein (10-fold). Any interrelationship between this TPA response
and the DNA-alkylation response was studied by treatment with
combinations of MNNG and TPA. The
-pol up-regulation observed with
MNNG treatment was abrogated by TPA, and conversely the up-regulation
observed with TPA treatment was abrogated by MNNG.
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