Translocation of a UV-damaged DNA binding protein into a tight association with chromatin after treatment of mammalian cells with UV light

Vesna Rapić Otrin, Mary McLenigan, Masashi Takao, Arthur S. Levine, Miroslava Protić

Research output: Contribution to journalArticlepeer-review

63 Citations (Scopus)

Abstract

A UV-damaged DNA binding protein (UV-DDB) is the major source of UV-damaged DNA binding activity in mammalian cell extracts. This activity is defective in at least some xeroderma pigmentosum group E (XP-E) patients; microinjection of the UV-DDB protein into their fibroblasts corrects nucleotide excision repair (NER). In an in vitro reconstituted NER system, small amounts of UV-DDB stimulate repair synthesis a few fold. After exposure to UV, mammalian cells show an early dose-dependent inhibition of the extractable UV-DDB activity; this inhibition may reflect a tight association of the binding protein with UV-damaged genomic DNA. To investigate the dynamics and location of UV-DDB with respect to damaged chromatin in vivo, we utilized nuclear fractionation and specific antibodies and detected translocation of the p127 component of UV-DDB from a loose to a tight association with chromatinized DNA immediately after UV treatment. A similar redistribution was found for other NER proteins, i.e. XPA, RP-A and PCNA, suggesting their tighter association with genomic DNA after UV. These studies revealed a specific protein-protein interaction between UV-DDB/p127 and RP-A that appears to enhance binding of both proteins to UV-damaged DNA in vitro, providing evidence for the involvement of UV-DDB in the damage-recognition step of NER. Moreover, the kinetics of the reappearance of extractable UV-DDB activity after UV treatment of human cells with differing repair capacities positively correlate with the cell's capacity to repair 6-4 pyrimidine dimers (6-4 PD) in the whole genome, a result consistent with an in vivo role for UV-DDB in recognizing this type of UV lesion.

Original languageEnglish
Pages (from-to)1159-1168
Number of pages10
JournalJournal of cell science
Volume110
Issue number10
Publication statusPublished - 1997 May 1

Keywords

  • Chromatin
  • DNA binding protein
  • DNA repair
  • UV light
  • Xeroderma pigmentosum

ASJC Scopus subject areas

  • Cell Biology

Fingerprint Dive into the research topics of 'Translocation of a UV-damaged DNA binding protein into a tight association with chromatin after treatment of mammalian cells with UV light'. Together they form a unique fingerprint.

Cite this