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PMID: 11564859 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The xeroderma pigmentosum group E gene product DDB2 is a specific target of cullin 4A in mammalian cells.

Molecular and cellular biology ·Vol. 21 ·No. 20 ·2001-10-00 ·Pages 6738-47

Nag A, Bondar T, Shiv S, Raychaudhuri P

Abstract

The damaged-DNA binding protein DDB consists of two subunits, DDB1 (127 kDa) and DDB2 (48 kDa). Mutations in the DDB2 subunit have been detected in patients suffering from the repair deficiency disease xeroderma pigmentosum (group E). In addition, recent studies suggested a role for DDB2 in global genomic repair. DDB2 also exhibits transcriptional activity. We showed that expression of DDB1 and DDB2 stimulated the activity of the cell cycle regulatory transcription factor E2F1. Here we show that DDB2 is a cell cycle-regulated protein. It is present at a low level in growth-arrested primary fibroblasts, and after release the level peaks at the G(1)/S boundary. The cell cycle regulation of DDB2 involves posttranscriptional mechanisms. Moreover, we find that an inhibitor of 26S proteasome increases the level of DDB2, suggesting that it is regulated by the ubiquitin-proteasome pathway. Our previous study indicated that the cullin family protein Cul-4A associates with the DDB2 subunit. Because cullins are involved in the ubiquitin-proteasome pathway, we investigated the role of Cul-4A in regulating DDB2. Here we show that DDB2 is a specific target of Cul-4A. Coexpression of Cul-4A, but not Cul-1 or other highly related cullins, increases the ubiquitination and the decay rate of DDB2. A naturally occurring mutant of DDB2 (2RO), which does not bind Cul-4A, is not affected by coexpression of Cul-4A. Studies presented here identify a specific function of the Cul-4A gene, which is amplified and overexpressed in breast cancers.

MeSH Terms
Animals Blotting, Western Cell Cycle Cell Cycle Proteins Cell Line Cell Nucleus/metabolism Cullin Proteins Cycloheximide/pharmacology DNA/metabolism DNA Repair DNA, Complementary/metabolism DNA-Binding Proteins/genetics,metabolism E2F Transcription Factors E2F1 Transcription Factor Fibroblasts/metabolism HeLa Cells Humans Mutation Neoplasm Proteins/metabolism Peptide Hydrolases Plasmids/metabolism Precipitin Tests Proteasome Endopeptidase Complex Protein Synthesis Inhibitors/pharmacology RNA Processing, Post-Transcriptional Ribonucleases/metabolism Time Factors Transcription Factors Transcription, Genetic Transfection Tumor Cells, Cultured Ubiquitins/pharmacology
Chemicals
CUL4A protein, human Cell Cycle Proteins Cullin Proteins DDB1 protein, human DDB2 protein, human DNA, Complementary DNA-Binding Proteins E2F Transcription Factors E2F1 Transcription Factor E2F1 protein, human Neoplasm Proteins Protein Synthesis Inhibitors Transcription Factors Ubiquitins DNA Cycloheximide Ribonucleases Peptide Hydrolases Proteasome Endopeptidase Complex ATP dependent 26S protease
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Nag A
Department of Biochemistry and Molecular Biology (M/C 536), University of Illinois at Chicago, Chicago, Illinois 60612, USA.
Bondar T
Shiv S
Raychaudhuri P
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.21.20.6738-6747.2001
Published
2001-10-00
Pages
6738-47
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC99852
Subset
IM
Grants
NCI NIH HHS · R01 CA077637 · United States
NCI NIH HHS · R01 CA088863 · United States
NCI NIH HHS · CA77637 · United States
NCI NIH HHS · CA88863 · United States
External Links
PubMed source DOI: 10.1128/MCB.21.20.6738-6747.2001 PMC Full Text View journal details
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