Enhancement of the in vitro transcription by T7 RNA polymerase of short DNA templates containing oxidative thymine lesions.
5-(Hydroxymethyl)uracil, 5-formyluracil or 5-carboxyuracil can be formed in DNA upon exposure to ionizing radiations. We report the effect of the latter oxidative lesions on the in vitro transcription by T7 RNA polymerase using a single-stranded short model template. Kinetic experiments showed that the transcription yield was increased with the modified template. Due to the unforeseen presence of a hairpin structure within the transcribed RNA, we postulated that we are in the presence of a T7 RNA polymerase pausing signal and that the lesions within this critical DNA structure could interfere with normal regulation of RNA transcription.
Guerniou
Valérie
V
Laboratoire Lésions des acides nucléiques, Service de chimie inorganique et biologique, FRE 2600, CEA/DSM/Département de recherche fondamentale sur la matière condensée, CEA-Grenoble, 17, rue des Martyrs, 38054 Grenoble cedex 9, France.
Gasparutto
Didier
D
Douki
Thierry
T
Cadet
Jean
J
Sauvaigo
Sylvie
S
D017123:Bacteriophage T7 / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D004249:DNA Damage; D017931:DNA Primers; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D007700:Kinetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D018384:Oxidative Stress; D015800:Protozoan Proteins; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D013698:Templates, Genetic; D013936:Thymidine / Q000031:analogs & derivatives*; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics*
C500541:D0295c protein, Plasmodium falciparum; D017931:DNA Primers; D015800:Protozoan Proteins; D012367:RNA, Viral; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D013936:Thymidine