title: 5'-derivatives of oligonucleotides as primers of DNA polymerization catalyzed by AMV reverse transcriptase and Klenow fragment of DNA polymerase 1.
abstract: The Km and Vmax values for d(pT)8 and its derivatives containing various 5'-end groups were estimated in the reaction of polymerization catalyzed with AMV-RT and FK. The change in affinity of modified primers was more pronounced in the case of AMV-RT than in the case of FK. Introducing in d(pT)8 of intercalators such as phenazinium, ethidium and daunomycin residues results in 2.7-, 8.7- and 11-fold increases in the primer affinity to AMV-RT, respectively. However, in the case of hemin and cholesterol derivatives the Km values were 3 and 5 times higher than those for d(pT)8. Compared to d(pT)8, the affinity of FK to all the above analogs was 2.3-3.6 times higher with the exception of cholesterol derivative to which it was 2.4-fold lower. The effect of the 5'-end residues on the Vmax values of d(pT)8 was small and ranged from 44% to 120% of that for d(pT)8. Therefore such reactive derivatives of oligonucleotides can be used as effective primers of AMV-RT and FK. Possible reasons for various effects of the 5'-end residues of the primer on its interaction with FK or AMV-RT in the presence of poly(A) are discussed.
authors.affiliation: Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of the Academy of Sciences of the USSR.
authors.forename: I A
authors.initials: IA
authors.lastname: Lokhova
authors.forename: G A
authors.initials: GA
authors.lastname: Nevinsky
authors.forename: T S
authors.initials: TS
authors.lastname: Godovikova
authors.forename: E M
authors.initials: EM
authors.lastname: Ivanova
authors.forename: A A
authors.initials: AA
authors.lastname: Koshkin
authors.forename: D S
authors.initials: DS
authors.lastname: Sergeev
authors.forename: E I
authors.initials: EI
authors.lastname: Frolova
authors.forename: N K
authors.initials: NK
authors.lastname: Rudenko
authors.forename: V V
authors.initials: VV
authors.lastname: Khomov
authors.forename: V M
authors.initials: VM
authors.lastname: Kavsan
mesh_terms: D009189:Avian Myeloblastosis Virus / Q000201:enzymology*; D004256:DNA Polymerase I / Q000378:metabolism*; D004261:DNA Replication; D007700:Kinetics; D015345:Oligonucleotide Probes / Q000138:chemical synthesis / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: D015345:Oligonucleotide Probes; D010446:Peptide Fragments; D012194:RNA-Directed DNA Polymerase; D004256:DNA Polymerase I