UV-specific DNA repair recombinant fusion enzyme: a new stable pharmacologically active principle suitable for photoprotection.
BACKGROUND
UV radiation can produce mutations in skin cells and correlates strongly with the onset of actinic keratoses and basal and squamous cell carcinomas. Xeroderma pigmentosum (XP) is a heritable disease characterized by an extreme sensitivity of skin to UV radiation. Recently, studies in cultured cells as well as in XP patients have demonstrated that the recombinant T4 endonuclease V UV-specific endonuclease could enhance repair of UV-induced photoproducts.


OBJECTIVE
We aimed to obtain a stable UV-specific DNA recombinant endonuclease, pharmacologically active in mammalian cells so as to be used in treatment and prophylaxis of sun damage.


METHODS
The UV-specific DNA endonuclease gene obtained from Micrococcus luteus, was fused to a leader peptide and expressed (alphaUveA), refolded and purified. A construction under the control of an eukaryotic promoter was used to transfect XP fibroblasts deficient in DNA damage repair. Transformed cells were UV irradiated and cell survival was assessed.


RESULTS
alphaUveA was obtained as a highly active UV-specific repair enzyme stable for at least 2 years. XP fibroblasts transfected with alphaUveA gene increased the resistance to UV radiation and, in consequence, cell survival.


CONCLUSIONS
alphaUveA is stable and pharmacologically active in human cells. The topical administration of this long-term stable new active principle could help diminish the risks of skin cancer after sun exposure.
Basílico
Guillermo
G
Laboratorio de Medicina Molecular, Fundación Pablo Cassará, Buenos Aires, Argentina.
Roger
Carolina Alvarez
CA
Seigelchifer
Mauricio
M
Kerner
Nestor
N
D002460:Cell Line; D002470:Cell Survival; D004247:DNA / Q000378:metabolism; D004249:DNA Damage; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000378:metabolism; D004260:DNA Repair; D043224:Deoxyribonuclease (Pyrimidine Dimer) / Q000378:metabolism; D004307:Dose-Response Relationship, Radiation; D004587:Electrophoresis, Agar Gel; D005347:Fibroblasts / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D008027:Light; D016982:Micrococcus luteus / Q000201:enzymology; D009369:Neoplasms / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013471:Sunburn; D013696:Temperature; D013997:Time Factors; D014162:Transfection; D014466:Ultraviolet Rays; D014764:Viral Proteins / Q000378:metabolism; D014983:Xeroderma Pigmentosum / Q000378:metabolism
D010455:Peptides; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D004247:DNA; C470698:endonuclease V, phage T4; D043224:Deoxyribonuclease (Pyrimidine Dimer); D011088:DNA Ligases; D000072481:DNA Ligase ATP