TY - JOUR
T1 - PCNA-induced DNA synthesis past cis-syn and trans-syn-l thymine dimers by calf thymus DNA polymerase δ in vitro
AU - O'day, Christine
AU - Burgers, Peter M.J.
AU - Taylor, John stephen
N1 - Funding Information:
This investigation was supported by PHS Grant number R01-CA40463 (to J.-S. T.) and GM32431 (to P.M.J.B) awarded by the National Institutes of Health.
PY - 1992/10/25
Y1 - 1992/10/25
N2 - Calf thymus proliferating cell nuclear antigen (PCNA) promoted DNA synthesis past cis-syn and trans-syn-l cyclobutane thymine dimers by calf thymus DNA polymerase δ (Polδ) in vitro. Templates containing site-specific cis-syn and trans-syn-l thymine dimers were prepared via a combination of solid phase synthesis with photoproduct building blocks and DNA lligation. Extension of a 15-mer primer on the UV dlmer-containing templates by Polδ produced termination and bypass products in a dNTP and PCNA dependent manner. In the absence of PCNA and at dNTP concentrations varying between 1 and 100 μM, Polδ could not bypass the cis-syn dlmer and terminated elongation one nucleotkle prior to the 3-T of the dimer. DNA synthesis past the trans-syn-l dimer was even less efficient. In the presence of PCNA, termination occurred primarily one nucleotlde prior to the 3′-T of both dimers at 1 μM dNTPs but opposite the 5′-T of the dimers at 100 μM dNTPs. In addition, under the latter conditions, bypass of the dimers was observed, to the extent of about 30% of the products for the cis-syn dimer and about 15% for the trans-syn-l dlmer.
AB - Calf thymus proliferating cell nuclear antigen (PCNA) promoted DNA synthesis past cis-syn and trans-syn-l cyclobutane thymine dimers by calf thymus DNA polymerase δ (Polδ) in vitro. Templates containing site-specific cis-syn and trans-syn-l thymine dimers were prepared via a combination of solid phase synthesis with photoproduct building blocks and DNA lligation. Extension of a 15-mer primer on the UV dlmer-containing templates by Polδ produced termination and bypass products in a dNTP and PCNA dependent manner. In the absence of PCNA and at dNTP concentrations varying between 1 and 100 μM, Polδ could not bypass the cis-syn dlmer and terminated elongation one nucleotkle prior to the 3-T of the dimer. DNA synthesis past the trans-syn-l dimer was even less efficient. In the presence of PCNA, termination occurred primarily one nucleotlde prior to the 3′-T of both dimers at 1 μM dNTPs but opposite the 5′-T of the dimers at 100 μM dNTPs. In addition, under the latter conditions, bypass of the dimers was observed, to the extent of about 30% of the products for the cis-syn dimer and about 15% for the trans-syn-l dlmer.
UR - http://www.scopus.com/inward/record.url?scp=0026715131&partnerID=8YFLogxK
U2 - 10.1093/nar/20.20.5403
DO - 10.1093/nar/20.20.5403
M3 - Article
C2 - 1359505
AN - SCOPUS:0026715131
VL - 20
SP - 5403
EP - 5406
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 0305-1048
IS - 20
ER -