ATP-dependent chromatin remodeling facilitates nucleotide excision repair of UV-induced DNA lesions in synthetic dinucleosomes.
To investigate the relationship between chromatin dynamics and nucleotide excision repair (NER), we have examined the effect of chromatin structure on the formation of two major classes of UV-induced DNA lesions in reconstituted dinucleosomes. Furthermore, we have developed a model chromatin-NER system consisting of purified human NER factors and dinucleosome substrates that contain pyrimidine (6-4) pyrimidone photoproducts (6-4PPs) either at the center of the nucleosome or in the linker DNA. We have found that the two classes of UV-induced DNA lesions are formed efficiently at every location on dinucleosomes in a manner similar to that of naked DNA, even in the presence of histone H1. On the other hand, excision of 6-4PPs is strongly inhibited by dinucleosome assembly, even within the linker DNA region. These results provide direct evidence that the human NER machinery requires a space greater than the size of the linker DNA to excise UV lesions efficiently. Interestingly, NER dual incision in dinucleosomes is facilitated by recombinant ACF, an ATP-dependent chromatin remodeling factor. Our results indicate that there is a functional connection between chromatin remodeling and the initiation step of NER.
Ura
K
K
Division of Gene Therapy Science, Osaka University School of Medicine, 2-2 Yamada-oka, Suita, Japan. kiyoeura@gts.med.osaka-u.ac.jp
Araki
M
M
Saeki
H
H
Masutani
C
C
Ito
T
T
Iwai
S
S
Mizukoshi
T
T
Kaneda
Y
Y
Hanaoka
F
F
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism*; D004249:DNA Damage; D004260:DNA Repair; D009707:Nucleosomes / Q000528:radiation effects*; D011740:Pyrimidine Dimers / Q000378:metabolism; D013379:Substrate Specificity; D014157:Transcription Factors / Q000378:metabolism*; D014466:Ultraviolet Rays / Q000009:adverse effects*
D002843:Chromatin; C086468:ISWI protein; D009707:Nucleosomes; D011740:Pyrimidine Dimers; D014157:Transcription Factors; C058406:pyrimidine-pyrimidone dimer; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases