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PMID: 16940174 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

DDB1 maintains genome integrity through regulation of Cdt1.

Molecular and cellular biology ·Vol. 26 ·No. 21 ·2006-11-00 ·Pages 7977-90

Lovejoy CA, Lock K, Yenamandra A, Cortez D

Abstract

DDB1, a component of a Cul4A ubiquitin ligase complex, promotes nucleotide excision repair (NER) and regulates DNA replication. We have investigated the role of human DDB1 in maintaining genome stability. DDB1-depleted cells accumulate DNA double-strand breaks in widely dispersed regions throughout the genome and have activated ATM and ATR cell cycle checkpoints. Depletion of Cul4A yields similar phenotypes, indicating that an E3 ligase function of DDB1 is important for genome maintenance. In contrast, depletion of DDB2, XPA, or XPC does not cause activation of DNA damage checkpoints, indicating that defects in NER are not involved. One substrate of DDB1-Cul4A that is crucial for preventing genome instability is Cdt1. DDB1-depleted cells exhibit increased levels of Cdt1 protein and rereplication, despite containing other Cdt1 regulatory mechanisms. The rereplication, accumulation of DNA damage, and activation of checkpoint responses in DDB1-depleted cells require entry into S phase and are partially, but not completely, suppressed by codepletion of Cdt1. Therefore, DDB1 prevents DNA lesions from accumulating in replicating human cells, in part by regulating Cdt1 degradation.

MeSH Terms
Animals Ataxia Telangiectasia Mutated Proteins Cell Cycle/physiology Cell Cycle Proteins/genetics,metabolism Cell Line Chromosomes, Human Cullin Proteins/metabolism DNA Damage DNA Repair DNA Replication DNA-Binding Proteins/genetics,metabolism Genes, cdc Genome, Human Genomic Instability Humans Protein Serine-Threonine Kinases/genetics,metabolism Protein Subunits/genetics,metabolism RNA, Small Interfering/genetics,metabolism S-Phase Kinase-Associated Proteins/genetics,metabolism Tumor Suppressor Proteins/genetics,metabolism Xeroderma Pigmentosum Xeroderma Pigmentosum Group A Protein/genetics,metabolism
Chemicals
CDT1 protein, human CUL4A protein, human Cell Cycle Proteins Cullin Proteins DDB1 protein, human DNA-Binding Proteins Protein Subunits RNA, Small Interfering S-Phase Kinase-Associated Proteins Tumor Suppressor Proteins XPA protein, human Xeroderma Pigmentosum Group A Protein XPC protein, human ATM protein, human ATR protein, human Ataxia Telangiectasia Mutated Proteins Protein Serine-Threonine Kinases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Lovejoy Courtney A
Department of Biochemistry, Vanderbilt University, Nashville, TN 37232, USA.
Lock Kimberli
Yenamandra Ashwini
Cortez David
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.00819-06
Published
2006-11-00
Epub
2006-00-28
Pages
7977-90
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC1636754
Subset
IM
Grants
NCI NIH HHS · R01 CA102729 · United States
External Links
PubMed source DOI: 10.1128/MCB.00819-06 PMC Full Text View journal details
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