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Nucleic Acids Research, Vol 24, Issue 8 1561-1565, Copyright © 1996 by Oxford University Press

ARTICLES

Mutation spectra of TA*, the major photoproduct of thymidylyl-(3'5')- deoxyadenosine, in Escherichia coli under SOS conditions

X Zhao and JS Taylor
Department of Chemistry, Washington University, St Louis, MO 63130- 4899, USA.

The biological activity of TA*, the major photoproduct of thymidylyl- (3',5')-deoxyadenosine, has remained speculative since it was identified a decade ago. To determine the mutagenicity of TA* in Escherichia coli, we constructed the replicative form of an M13mp18- derived phage containing TA* in the (-)-strand by polymerase-catalyzed elongation of a TA*-containing 49mer opposite a uracil-containing (+)- strand of the phage. The in vitro synthesis mixture was transfected into an ung+, phr- E.coli host and the progeny were screened with a hybridization probe unique for the (-)-strand. TA* was found to block DNA replication substantially in the absence of SOS, but under SOS, TA* was bypassed more efficiently and was highly mutagenic. Among 56 analyzed (-)-strand progeny from two transfections, 46 (82%) were mutants, including six (11%) tandem mutants. The most abundant mutation was a 3'A-->T substitution (31/46, 56%). The possible biological consequences of TA* formation in the highly conserved TATA box consensus sequence on gene expression are discussed in light of the mutagenicity of TA*.
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