Chromatin structure modulates DNA repair by photolyase in vivo.
Yeast and many other organisms use nucleotide excision repair (NER) and photolyase in the presence of light (photoreactivation) to repair cyclobutane pyrimidine dimers (CPDs), a major class of DNA lesions generated by UV light. To study the role of photoreactivation at the chromatin level in vivo, we used yeast strains which contained minichromosomes (YRpTRURAP, YRpCS1) with well-characterized chromatin structures. The strains were either proficient (RAD1) or deficient (rad1 delta) in NER. In contrast to NER, photolyase rapidly repairs CPDs in non-nucleosomal regions, including promoters of active genes (URA3, HIS3, DED1) and in linker DNA between nucleosomes. CPDs in nucleosomes are much more resistant to photoreactivation. These results demonstrate a direct role of chromatin in modulation of a DNA repair process and an important role of photolyase in repair of damaged promoters with presumptive effects on gene regulation. In addition, photoreactivation provides an in vivo test for chromatin structure and stability. In active genes (URA3, HIS3), photolyase repairs the non-transcribed strand faster than the transcribed strand and can match fast removal of lesions from the transcribed strand by NER (transcription-coupled repair). Thus, the combination of both repair pathways ensures efficient repair of active genes.
Suter
B
B
Institut für Zellbiologie, ETH-Hönggerberg, Zürich, Switzerland.
Livingstone-Zatchej
M
M
Thoma
F
F
D002843:Chromatin / Q000737:chemistry*; D015825:Chromosomes, Fungal; D004249:DNA Damage; D004260:DNA Repair / Q000502:physiology* / Q000528:radiation effects; D045643:DNA Repair Enzymes; D004268:DNA-Binding Proteins; D004255:Deoxyribodipyrimidine Photo-Lyase / Q000378:metabolism*; D004720:Endonucleases / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D008027:Light; D008957:Models, Genetic; D009154:Mutation; D009707:Nucleosomes; D011401:Promoter Regions, Genetic; D011740:Pyrimidine Dimers; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000528:radiation effects; D029701:Saccharomyces cerevisiae Proteins; D014466:Ultraviolet Rays
D002843:Chromatin; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D009707:Nucleosomes; D011740:Pyrimidine Dimers; D029701:Saccharomyces cerevisiae Proteins; D004720:Endonucleases; C069266:RAD1 protein, S cerevisiae; D004255:Deoxyribodipyrimidine Photo-Lyase; D045643:DNA Repair Enzymes