3,N(4)-ethano-2'-deoxycytidine: chemistry of incorporation into oligomeric DNA and reassessment of miscoding potential.
3,N(4)-Ethano-2'-deoxycytidine (ethano-dC) may be incorporated successfully into synthetic oligodeoxynucleotides by omitting the capping procedure used in the automated DNA synthetic protocols immediately after inserting the lesion and in all iterations thereafter. Ethano-dC is sensitive to acetic anhydride found in the capping reagent, and multiple oligomeric products are formed. These products were identified by examining the reaction of ethano-dC with the capping reagent, and several acetylated, ring-opened products were characterized by electrospray mass spectrometry and collision induced dissociation experiments on a tandem quadrupole mass spectrometer. A scheme for the formation of the acetylated products is proposed. In addition, the mutagenic profile of ethano-dC was re-examined and compared to that for etheno-dC. Ethano-dC is principally a blocking lesion; however, when encountered by the exo(-)Klenow fragment of DNA polymerase, dAMP (22%), TMP (16%), dGMP (5.3%) and dCMP (1.2%) were all incorporated opposite ethano-dC, along with an oligomer containing a one-base deletion (0.6%).
Bonala
R R
RR
Department of Pharmacological Sciences, State University of New York at Stony Brook, Stony Brook, NY 11794-3400, USA.
Rieger
R A
RA
Shibutani
S
S
Grollman
A P
AP
Iden
C R
CR
Johnson
F
F
D000107:Acetylation; D001331:Automation; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000138:chemical synthesis* / Q000737:chemistry; D004256:DNA Polymerase I / Q000378:metabolism; D003841:Deoxycytidine / Q000031:analogs & derivatives / Q000494:pharmacology; D007202:Indicators and Reagents; D015394:Molecular Structure; D009153:Mutagens / Q000737:chemistry / Q000494:pharmacology; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis* / Q000737:chemistry*; D018629:Spectrometry, Mass, Secondary Ion
D007202:Indicators and Reagents; D009153:Mutagens; D009838:Oligodeoxyribonucleotides; D003841:Deoxycytidine; C032222:3,N(4)-ethenodeoxycytidine; D004247:DNA; D004256:DNA Polymerase I