Df. Kusewitt et al., ENHANCED PYRIMIDINE DIMER REPAIR IN CULTURED MURINE EPITHELIAL-CELLS TRANSFECTED WITH THE DENV GENE OF BACTERIOPHAGE-T4, Journal of investigative dermatology, 102(4), 1994, pp. 485-489
The patch size for excision repair of ultraviolet radiation (UV)-induc
ed pyrimidine dimers was determined in cultured murine epithelial cell
s with normal and enhanced pyrimidine dimer repair capabilities. Cells
with enhanced pyrimidine dimer repair were produced by transfecting 3
08 cells with the denV gene of bacteriophage T4; this gene encodes the
enzyme endonuclease V. Pyrimidine dimer repair following exposure to
UV from an FS-40 sunlamp was determined by micrococcal dimer-specific
nuclease digestion and alkaline sucrose ultracentrifugation. Patch siz
e was estimated based on the photolytic lability of bromodeoxyuridine-
substituted DNA. Excision repair or UV-induced pyrimidine dimers in de
nV-transfected 308 cells was enhanced two- to threefold. Production of
mRNA from the denV gene in cell lines with enhanced repair was confir
med by RNA blotting. In control cells, the patch size for excision rep
air of DNA photoproducts was estimated to be 34 nucleotides per photop
roduct removed; in denV-transfected cells, a smaller average patch siz
e of 10-16 nucleotides per photoproduct removed was calculated. Thus,
endonuclease V activity appears to alter not only the extent, but also
the nature of excision repair in UV-exposed mammalian epithelial cell
s.