Synthesis and base pairing properties of DNA-RNA heteroduplex containing 5-hydroxyuridine.
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 approximately 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.
Cui
Song
S
Department of Biochemistry, College of Natural Sciences, Kyungpook National University, Daegu 702-701, Korea. cuisongpop@hanmail.net
Kim
Yong-Hoon
YH
Jin
Cheng-Hao
CH
Kim
Sang Kook
SK
Rhee
Man-Hee
MH
Kwon
Oh-Shin
OS
Moon
Byung Jo
BJ
D020029:Base Pairing / Q000502:physiology*; D002942:Circular Dichroism; D004247:DNA / Q000737:chemistry*; D008954:Models, Biological; D009691:Nucleic Acid Denaturation / Q000528:radiation effects; D009692:Nucleic Acid Heteroduplexes / Q000138:chemical synthesis* / Q000737:chemistry; D012313:RNA / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet; D014529:Uridine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology
D009692:Nucleic Acid Heteroduplexes; D012313:RNA; D004247:DNA; C010194:5-hydroxyuridine; D014529:Uridine