Premium
Effect of P‐chirality of oligo(deoxyribonucleoside phosphorothioate)s on the activity of terminal deoxyribonucleotidyl transferase
Author(s) -
Koziołkiewicz Maria,
Maciaszek Anna,
Stec Wojciech J,
Semizarov Dmitry,
Victorova Lyubov,
Krayevsky Alexander
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00900-4
Subject(s) - primer (cosmetics) , transferase , oligonucleotide , elongation , chemistry , stereochemistry , nucleotide , nucleoside , enzyme , microbiology and biotechnology , biochemistry , dna , biology , gene , organic chemistry , materials science , metallurgy , ultimate tensile strength
Phosphorothioate analogues of oligonucleotides (PS‐oligos) of predetermined chirality at the phosphorus atom at each internucleotide linkage have been used as primers for terminal deoxyribonucleotidyl transferase ( TdT , EC 2.7.7.31). The enzyme catalyzes efficient elongation of PS primers in which all phosphorothioate internucleotide linkages are uniformly of the [R P ] configuration, while the presence of the linkage(s) of the [S P ] configuration significantly decreases or completely inhibits the primer extension. Our results indicate that for the elongation of phosphorothioate oligomers the most important is the internucleotide bond located between the second and the third nucleoside from the 3′‐end. The presence of [S P ] linkage at this position strongly reduces the enzyme activity while the [R P ] bond allows for effective elongation of the primer. The activity of the enzyme is also influenced by base composition and sequence of phosphorothioate primer as well as the dNTP used for elongation process.