Effects of 8-chlorodeoxyadenosine on DNA synthesis by the Klenow fragment of DNA polymerase I.
8-chloro-2'-deoxyadenosine (8-Cl-dAdo) was incorporated into synthetic DNA oligonucleotides to determine its effects on DNA synthesis by the 3'-5' exonuclease-free Klenow fragment of Escherichia coli DNA Polymerase I (KF-). Single nucleotide insertion experiments were used to determine the coding potential of 8-Cl-dAdo in a DNA template. KF- inserted TTP opposite 8-Cl-dAdo in the template, but with decreased efficiency relative to natural deoxyadenosine. Running-start primer extensions with KF- resulted in polymerase pausing at 8-Cl-dAdo template sites during DNA synthesis. The 2'-deoxyribonucleoside triphosphate analogue, 8-Cl-dATP, was incorporated opposite thymidine (T) approximately two-fold less efficiently than dATP.
Chen
Lisa S
LS
Department of Chemistry, Robert H. Lurie Comprehensive Cancer Center, Northwestern University, 2145 Sheridan Road, Evanston, IL 60208-3113, USA.
Bahr
Michael H
MH
Sheppard
Terry L
TL
D004247:DNA / Q000138:chemical synthesis*; D004256:DNA Polymerase I / Q000737:chemistry*; D003839:Deoxyadenosines / Q000494:pharmacology*; D004926:Escherichia coli / Q000737:chemistry; D009841:Oligonucleotides / Q000737:chemistry*; D013936:Thymidine / Q000737:chemistry
C479978:8-chlorodeoxyadenosine; D003839:Deoxyadenosines; D009841:Oligonucleotides; D004247:DNA; D004256:DNA Polymerase I; D013936:Thymidine