Polymerase incorporation of a 2'-deoxynucleoside-5'-triphosphate bearing a 4-hydroxy-2-mercaptobenzimidazole nucleobase analogue

Kunihiko Morihiro; Hidekazu Hoshino; Osamu Hasegawa; Yuuya Kasahara; Kohsuke Nakajima; Masayasu Kuwahara; Shin-ichi Tsunoda; Satoshi Obika

doi:10.1016/j.bmcl.2015.05.075

Polymerase incorporation of a 2'-deoxynucleoside-5'-triphosphate bearing a 4-hydroxy-2-mercaptobenzimidazole nucleobase analogue

Bioorg Med Chem Lett. 2015 Aug 1;25(15):2888-91. doi: 10.1016/j.bmcl.2015.05.075. Epub 2015 May 29.

Authors

Kunihiko Morihiro¹, Hidekazu Hoshino², Osamu Hasegawa², Yuuya Kasahara³, Kohsuke Nakajima⁴, Masayasu Kuwahara⁴, Shin-ichi Tsunoda⁵, Satoshi Obika⁶

Affiliations

¹ National Institute of Biomedical Innovation, Health and Nutrition (NIBIOHN), 7-6-8 Saito-Asagi, Ibaraki, Osaka 567-0085, Japan; Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: k-morihiro@nibiohn.go.jp.
² Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
³ National Institute of Biomedical Innovation, Health and Nutrition (NIBIOHN), 7-6-8 Saito-Asagi, Ibaraki, Osaka 567-0085, Japan; Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan.
⁴ Graduate School of Science and Technology, Gunma University, 1-5-1 Tenjin-Cho, Kiryu, Gunma 376-8515, Japan.
⁵ National Institute of Biomedical Innovation, Health and Nutrition (NIBIOHN), 7-6-8 Saito-Asagi, Ibaraki, Osaka 567-0085, Japan.
⁶ National Institute of Biomedical Innovation, Health and Nutrition (NIBIOHN), 7-6-8 Saito-Asagi, Ibaraki, Osaka 567-0085, Japan; Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan. Electronic address: obika@phs.osaka-u.ac.jp.

PMID: 26048797
DOI: 10.1016/j.bmcl.2015.05.075

Abstract

Here, we describe the enzymatic construction of a new larger base pair formed between adenine (A) and a 4-hydroxy-2-mercaptobenzimidazole (SB) nucleobase analogue. We investigated the enzymatic incorporation of 2'-deoxynucleoside-5'-triphosphate bearing a SB nucleobase analogue (dSBTP) into oligonucleotides (ONs) by DNA polymerases. dSBTP could be effectively incorporated at the site opposite a dA in a DNA template by several B family DNA polymerases. These findings provide new insights into various aspects of biotechnology, including the design of non-natural base pairs.

Keywords: 5′-Triphosphates; DNA polymerases; Glycosidic conformation; Nucleobase modification; Primer extension.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenine / chemistry
Adenine / metabolism*
Base Pairing
Base Sequence
Benzimidazoles / chemistry
Benzimidazoles / metabolism*
DNA Primers / chemistry
DNA Primers / metabolism
DNA-Directed DNA Polymerase / metabolism*
Nucleotides / chemistry
Nucleotides / metabolism*
Oligonucleotides / chemistry
Oligonucleotides / metabolism
Polymerization
Polyphosphates / chemistry
Polyphosphates / metabolism*

Substances

Benzimidazoles
DNA Primers
Nucleotides
Oligonucleotides
Polyphosphates
2-mercaptobenzimidazole
DNA-Directed DNA Polymerase
Adenine
triphosphoric acid