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PMID: 16473935 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The DDB1-CUL4ADDB2 ubiquitin ligase is deficient in xeroderma pigmentosum group E and targets histone H2A at UV-damaged DNA sites.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 103 ·No. 8 ·2006-02-21 ·Pages 2588-93

Kapetanaki MG, Guerrero-Santoro J, Bisi DC, Hsieh CL, Rapić-Otrin V, Levine AS

Abstract

Xeroderma pigmentosum (XP) is a heritable human disorder characterized by defects in nucleotide excision repair (NER) and the development of skin cancer. Cells from XP group E (XP-E) patients have a defect in the UV-damaged DNA-binding protein complex (UV-DDB), involved in the damage recognition step of NER. UV-DDB comprises two subunits, products of the DDB1 and DDB2 genes, respectively. Mutations in the DDB2 gene account for the underlying defect in XP-E. The UV-DDB complex is a component of the newly identified cullin 4A-based ubiquitin E3 ligase, DDB1-CUL4A(DDB2). The E3 ubiquitin ligases recognize specific substrates and mediate their ubiquitination to regulate protein activity or target proteins for degradation by the proteasomal pathway. In this study, we have addressed the role of the UV-DDB-based E3 in NER and sought a physiological substrate. We demonstrate that monoubiquitinated histone H2A in native chromatin coimmunoprecipitates with the endogenous DDB1-CUL4A(DDB2) complex in response to UV irradiation. Further, mutations in DDB2 alter the formation and binding activity of the DDB1-CUL4A(DDB2) ligase, accompanied by impaired monoubiquitination of H2A after UV treatment of XP-E cells, compared with repair-proficient cells. This finding indicates that DDB2, as the substrate receptor of the DDB1-CUL4A-based ligase, specifically targets histone H2A for monoubiquitination in a photolesion-binding-dependent manner. Given that the loss of monoubiquitinated histone H2A at the sites of UV-damaged DNA is associated with decreased global genome repair in XP-E cells, this study suggests that histone modification, mediated by the XPE factor, facilitates the initiation of NER.

MeSH Terms
Chromatin/metabolism Cullin Proteins/analysis,metabolism DNA/chemistry,radiation effects DNA Damage DNA-Binding Proteins/analysis,genetics,metabolism Histones/analysis,metabolism Humans Mutation Tumor Cells, Cultured Ubiquitin Ubiquitin-Protein Ligases/analysis,metabolism Ultraviolet Rays Xeroderma Pigmentosum/enzymology,genetics
Chemicals
CUL4A protein, human Chromatin Cullin Proteins DDB1 protein, human DDB2 protein, human DNA-Binding Proteins Histones Ubiquitin DNA Ubiquitin-Protein Ligases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Kapetanaki Maria G
Department of Molecular Genetics and Biochemistry, School of Medicine, and Cancer Institute, University of Pittsburgh, Hillman Cancer Center, Research Pavilion, Suite 2.6, 5117 Centre Avenue, Pittsburgh, PA 15213, USA.
Guerrero-Santoro Jennifer
Bisi Dawn C
Hsieh Ching L
Rapić-Otrin Vesna
Levine Arthur S
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0511160103
Published
2006-02-21
Epub
2006-00-10
Pages
2588-93
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC1413840
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.0511160103 PMC Full Text View journal details
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