Nucleic Acids Research, Vol 24, Issue 10 1837-1840, Copyright © 1996 by Oxford University Press
A Gentil, F Le Page, A Margot, CW Lawrence, A Borden and A Sarasin
The mutagenic properties of UV-induced photoproducts, both the cis-syn
thymine-thymine dimer (TT) and the thymine-thymine pyrimidine pyrimidone
(6-4) photoproduct [T(6-4)T] were studied in mammalian cells using shuttle
vectors. A shuttle vector able to replicate in both mammalian cells and
bacteria was produced in its single-stranded DNA form. A unique
photoproduct was inserted at a single restriction site and after
recircularization of the single-stranded DNA vector, this latter was
transfected into simian COS7 cells. After DNA replication the vector was
extracted from cells and used to transform bacteria. Amplified DNA was
finally analyzed without any selective screening, DNA from randomly picked
bacterial colonies being directly sequenced. Our results show clearly that
both lesions are mutagenic, but at different levels. Mutation frequencies
of 2 and 60% respectively were observed with the TT dimer and the T(6-4)T.
With the TT dimer the mutations were targeted on the 3'-T. With the T(6-4)T
a large variety of mutations were observed. A majority of G-->T
transversions were semi-targeted to the base before the 5'-T of the
photoproduct. These kinds of mutations were not observed when the same
plasmid was transfected directly into SOS-induced JM105 bacteria or when
the T(6-4)T oligonucleotide inserted in a different plasmid was replicated
in SOS-induced SMH10 Escherichia coil bacteria. These semi-targeted
mutations are therefore the specific result of bypass of the T(6-4)T lesion
in COS7 cells by one of the eukaryotic DNA polymerases.
ARTICLES
Mutagenicity of a unique thymine-thymine dimer or thymine-thymine pyrimidine pyrimidone (6-4) photoproduct in mammalian cells
Institut de Recherches sur le Cancer, CNRS, IFC 1, Laboratory of Molecular Genetics, Villejuif, France.
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