title: The substrate activity of (S)-9-[3-hydroxy-(2-phosphonomethoxy)propyl]adenine diphosphate toward DNA polymerases alpha, delta and epsilon.
abstract: In this study, we examined the substrate potency of (S)-9-[3-hydroxy-(2-phosphonomethoxy)propyl]- adenine diphosphate (HPMPApp) toward DNA polymerases alpha, delta and epsilon. The efficiency of HPMPApp incorporation decreased in the order pol epsilon >pol delta =pol alpha and was from 5.4- to 23-fold lower than that of dATP. Depending on which template-primer was used, the HPMPAppKm value was 3.3- and 8.3- (pol alpha), 3- and 0.82- (pol delta) or 2-fold (pol epsilon) higher than dATPKm. The ability of HPMPA to accumulate in DNA decreased in the order pol epsilon >pol alpha >pol delta. The difference between the elongation rate of DNA with and without one HPMPA molecule at 3' termini was about 1.1-2.3 fold. The 3'-5'-exonuclease activity of pol delta and epsilon can excise HPMPA from DNA. These observations indicate that interaction of HPMPApp with pol alpha, delta and epsilon may contribute to its cellular toxicity and explain its antiviral activity against polyomavirus.
authors.affiliation: Institute of Organic Chemistry & Biochemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Birkus
authors.forename: Dominik
authors.initials: D
authors.lastname: Rejman
authors.forename: Miroslav
authors.initials: M
authors.lastname: Otmar
authors.forename: Ivan
authors.initials: I
authors.lastname: Votruba
authors.forename: Ivan
authors.initials: I
authors.lastname: Rosenberg
authors.forename: Antonin
authors.initials: A
authors.lastname: Holy
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D011756:Diphosphates / Q000737:chemistry / Q000378:metabolism*; D005092:Exonucleases / Q000378:metabolism; D007700:Kinetics; D015394:Molecular Structure; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry; D063065:Organophosphonates; D009943:Organophosphorus Compounds / Q000737:chemistry / Q000378:metabolism*; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D051381:Rats; D013379:Substrate Specificity; D013698:Templates, Genetic
chemical_list: D011756:Diphosphates; D009841:Oligonucleotides; D063065:Organophosphonates; D009943:Organophosphorus Compounds; D018809:Proliferating Cell Nuclear Antigen; D004247:DNA; C050127:9-(S)-(3-hydroxy-2-(phosphonomethoxy)propyl)adenine; D004259:DNA-Directed DNA Polymerase; D005092:Exonucleases; D000225:Adenine