Site-specific repair of cyclobutane pyrimidine dimers in a positioned nucleosome by photolyase and T4 endonuclease V in vitro

The EMBO Journal
U Schieferstein, F Thoma

Abstract

Since genomic DNA is folded into nucleosomes, and DNA damage is generated all over the genome, a central question is how DNA repair enzymes access DNA lesions and how they cope with nucleosomes. To investigate this topic, we used a reconstituted nucleosome (HISAT nucleosome) as a substrate to generate DNA lesions by UV light (cyclobutane pyrimidine dimers, CPDs), and DNA photolyase and T4 endonuclease V (T4-endoV) as repair enzymes. The HISAT nucleosome is positioned precisely and contains a long polypyrimidine region which allows one to monitor formation and repair of CPDs over three helical turns. Repair by photolyase and T4-endoV was inefficient in nucleosomes compared with repair in naked DNA. However, both enzymes showed a pronounced site-specific modulation of repair on the nucleosome surface. Removal of the histone tails did not substantially enhance repair efficiency nor alter the site specificity of repair. Although photolyase and T4-endoV are different enzymes with different mechanisms, they exhibited a similar site specificity in nucleosomes. This implies that the nucleosome structure has a decisive role in DNA repair by exerting a strong constraint on damage accessibility. These findings may serve as a model for dam...Continue Reading

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Citations

Sep 3, 1999·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·M Meijer, M J Smerdon
Sep 5, 2002·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Vincent van den BoomWim Vermeulen
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Jun 8, 2002·Journal of Molecular Biology·Bernhard Suter, Fritz Thoma
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Jan 19, 2002·EMBO Reports·Catherine M Green, Geneviève Almouzni
Nov 2, 2002·The EMBO Journal·Hilde NilsenAlain Verreault
Jul 30, 2004·Molecular and Cellular Biology·Christoph CapiaghiFritz Thoma
Feb 24, 2009·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Maxime TremblayAntonio Conconi
Jun 17, 2006·The EMBO Journal·Andrea BucceriFritz Thoma
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Sep 7, 2021·The EMBO Journal·Smitha SivapragasamJohn J Wyrick

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