Synthesis and base pairing properties of DNA-RNA heteroduplex containing 5-hydroxyuridine

被引：6

作者：

Cui, Song ^{[1
,2
]}

Kim, Yong-Hoon ^{[1
,3
]}

Jin, Cheng-Hao ^{[4
]}

Kim, Sang Kook ^{[1
]}

Rhee, Man-hee ^{[2
]}

Kwon, Oh-Shin ^{[1
]}

Moon, Byung Jo ^{[1
]}

机构：

[1] Kyungpook Natl Univ, Dept Biochem, Coll Nat Sci, Taegu 702701, South Korea

[2] Kyungpook Natl Univ, Lab Vet Physiol & Signaling, Coll Vet Med, Taegu 702701, South Korea

[3] Kyungpook Natl Univ, Coll Med, N&B Co Ltd, Taegu 702832, South Korea

[4] Sungkyunkwan Univ, Drug Targeting Lab, Coll Pharm, Seoul, South Korea

来源：

BMB REPORTS | 2009年 / 42卷 / 06期

关键词：

Base pairing; CD spectra; Melting temperature; Oligoribonucleotide; 5-Hydroxyuridine; MP2 THEORY LEVELS; BSSE-FREE DFT; THERMODYNAMIC STABILITY; REVERSE TRANSCRIPTASES; OXIDATIVE STRESS; CDNA SYNTHESIS; DAMAGE; 8-OXO-7,8-DIHYDROGUANOSINE; INSERTION; OLIGORIBONUCLEOTIDES;

D O I：

10.5483/BMBRep.2009.42.6.373

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

070307 [化学生物学]; 071010 [生物化学与分子生物学];

摘要：

5-Hydroxyuridine (5-OHU) is a major lesion of uridine and cytosine produced in RNA by various chemical oxidants. To elucidate its biochemical and biophysical effects on RNA replication, the site-specifically modified oligoribonucleotides containing 5-OHU were synthesized with C5-hydroxy-5'-O-DMTr-2'-TBDMS-uridine phosphoramidite using automated solid phase synthesis. The base-pairing properties of nucleotides opposite 5-OHU in 24 mer oligoribonulcleotides with dNTP were studied using three reverse transcriptases (Super-Script (TM) II-, AMV-, MMLV-RT) in cDNA synthesis. Adenine as well as guanine was incorporated preferentially by all reverse transcriptases. In the UV-melting temperature experiment, the results from the relative stabilities of the base pairs were A : 5-OHU > G : 5-OHU > T : 5-OHU approximate to C : 5-OHU. Circular Dichroism (CD) studies showed that DNA-RNA containing 5-OHU heteroduplexes exhibit a similar conformation between the A-type RNA and B-type DNA. These results suggest that 5-OHU from oxidative damage was mainly influenced by adenine mismatch. [BMB reports 2009; 42(6): 373-379]

引用

页码：373 / 379

页数：7

共 31 条

[1]

A comparative study of the thermal-stability of oligodeoxyribonucleotides containing 5-substituted 2′deoxyuridines [J].