Template-primer activity of 5-(hydroxymethyl)uracil-containing DNA for prokaryotic and eukaryotic DNA and RNA polymerases.
We have utilized Bacillus subtilis phage SPO-1 DNA as a model of irradiated DNA. In this phage, all thymine (Thy) residues are replaced by 5-(hydroxymethyl)uracil (5HmUra), which is a known irradiation-induced derivative of DNA Thy. SPO-1 phage is naturally devoid of other such irradiation-induced DNA lesions. DNase I activated SPO-1 phage DNA served as well as, or even better than, the control DNAs (Bacillus subtilis DNA and calf thymus DNA) as a template-primer for Escherichia coli, Micrococcus luteus, and human HL-60 cell DNA polymerases. Furthermore, the template activity of SPO-1 phage DNA was also superior when transcription with E. coli RNA polymerase was investigated. The results reported here indicated that the replacement of Thy by 5HmUra is not deleterious to template and primer functions during DNA or RNA synthesis.
Herrala
A M
AM
Laboratory of Molecular Hematology, University of Oulu, Finland.
Vilpo
J A
JA
D001412:Bacillus subtilis; D001435:Bacteriophages / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics* / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D006801:Humans; D007938:Leukemia; D008837:Micrococcus / Q000201:enzymology; D010432:Pentoxyl / Q000031:analogs & derivatives* / Q000378:metabolism; D013698:Templates, Genetic; D013941:Thymine; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D014498:Uracil / Q000031:analogs & derivatives*
D004279:DNA, Viral; C008055:5-hydroxymethyluracil; D014498:Uracil; D010432:Pentoxyl; D012321:DNA-Directed RNA Polymerases; D004259:DNA-Directed DNA Polymerase; D013941:Thymine