Nucleic Acids Research, Vol 26, Issue 11 2611-2617, Copyright © 1998 by Oxford University Press
H Kamiya, S Iwai and H Kasai
Two major ultraviolet-induced photolesions of TpT, a (6-4) photoproduct
[T(6-4)T] and a cis-syn cyclobutane TT dimer (T=T), were incorporated into
a predetermined site of one of the leading and lagging template strands of
a double-stranded vector, and the modified DNAs were transfected into
simian COS-7 cells. The DNAs replicated in the cells were recovered and
were transfected again into Escherichia coli. The DNA replication
efficiencies of plasmids containing T(6-4)T and T=T in the template strand
for lagging strand synthesis were 93 and 79%, respectively, as compared
with the unmodified DNA. Similar inhibitory effects were observed in the
cases of the photoproducts in the template strand for leading strand
synthesis (71 and 58%, respectively). These results indicated that T(6-4)T
blocked DNA replication more weakly than T=T during leading and lagging
strand syntheses in mammalian cells. The mutation frequencies of T(6-4)T
were 2.3 and 4.7% in the leading and lagging template strands,
respectively. The T=T lesion was less mutagenic and induced mutations with
0.2-0.7% frequencies. The T(6-4)T lesion primarily elicited 3'-T-->C
substitutions, and T=T induced various types of mutations. These results
indicate that T(6-4)T is more mutagenic than T=T during leading and lagging
strand syntheses in simian cells. Moreover, this is the first evidence that
shows T(6-4)T mainly elicits targeted substitutions at its 3'-T site in
mammalian cells.
ARTICLES
The (6-4) photoproduct of thymine-thymine induces targeted substitution mutations in mammalian cells
Department of Environmental Oncology, Institute of Industrial Ecological Sciences, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan. hirokam@med.uoeh-u.ac.jp
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