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Nucleic Acids Research, 2002, Vol. 30, No. 11 2588-2598
© 2002 Oxford University Press

Sequential binding of UV DNA damage binding factor and degradation of the p48 subunit as early events after UV irradiation

Vesna Rapic-Otrin, Mary P. McLenigan1, Dawn C. Bisi, Martin Gonzalez2 and Arthur S. Levine*

Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, E1240 BST, Pittsburgh, PA 15261, USA, 1Section on DNA Replication, Repair and Mutagenesis, National Institute of Child Health and Human Development, NIH, Bethesda, MD 20892-2725, USA and 2Department of Biology, University of Colorado-Denver, Denver, CO 80217-3364, USA

The UV-damaged DNA binding protein complex (UV-DDB) is implicated in global genomic nucleotide excision repair (NER) in mammalian cells. The complex consists of a heterodimer of p127 and p48. UV-DDB is defective in one complementation group (XP-E) of the heritable, skin cancer-prone disorder xeroderma pigmentosum. Upon UV irradiation of primate cells, UV-DDB associates tightly with chromatin, concomitant with the loss of extractable binding activity. We report here that an early event after UV, but not ionizing, radiation is the transient dose-dependent degradation of the small subunit, p48. Treatment of human cells with the proteasomal inhibitor NIP-L3VS blocks this UV-induced degradation of p48. In XP-E cell lines with impaired UV-DDB binding, p48 is resistant to degradation. UV-mediated degradation of p48 occurs independently of the expression of p53 and the cell’s proficiency for NER, but recovery of p48 levels at later times (12 h and thereafter) is dependent upon the capacity of the cell to repair non-transcribed DNA. In addition, we find that the p127 subunit of UV-DDB binds in vivo to p300, a histone acetyltransferase. The data support a functional connection between UV-DDB binding activity, proteasomal degradation of p48 and chromatin remodeling during early steps of NER.

* To whom correspondence should be addressed. Tel: +1 412 648 8977; Fax: +1 412 648 1236; Email: alevine{at}hs.pitt.edu


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