In vitro chromatin templates to study nucleotide excision repair

Research output: Contribution to journalArticlepeer-review

7 Scopus citations


In eukaryotic cells, DNA associates with histones and exists in the form of a chromatin hierarchy. Thus, it is generally believed that many eukaryotic cellular DNA processing events such as replication, transcription, recombination and DNA repair are influenced by the packaging of DNA into chromatin. This mini-review covers the current knowledge of DNA damage and repair in chromatin based on in vitro studies. Specifically, nucleosome assembly affects DNA damage formation in both random sequences and sequences with strong nucleosome-positioning signals such as 5S rDNA. At least three systems have been used to analyze the effect of nucleosome folding on nucleotide excision repair (NER) in vitro: (a) human cell extracts that have to rely on labeling of repair synthesis to monitor DNA repair, due to very low repair efficacy; (b) Xenopus oocyte nuclear extracts, that have very robust DNA repair efficacy, have been utilized to follow direct removal of DNA damage, (c) six purified human DNA repair factors (RPA, XPA, XPC, TFIIH, XPG, and XPF-ERCC1) that have been used to reconstitute excision repair in vitro. In general, the results have shown that nucleosome folding inhibits NER and, therefore, its activity must be enhanced by chromatin remodeling factors like SWI/SNF. In addition, binding of transcription factors such as TFIIIA to the 5S rDNA promoter also modulates NER efficacy.

Original languageEnglish
Pages (from-to)68-76
Number of pages9
JournalDNA Repair
StatePublished - 2015

Bibliographical note

Publisher Copyright:
© 2015 Elsevier B.V.


  • 5S rDNA
  • Chromatin
  • Chromatin remodeling
  • DNA damage
  • In vitro
  • Nucleotide excision repair

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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