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PMID: 19704162 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Functional interaction between the Fanconi Anemia D2 protein and proliferating cell nuclear antigen (PCNA) via a conserved putative PCNA interaction motif.

The Journal of biological chemistry ·Vol. 284 ·No. 42 ·2009-10-16 ·Pages 28935-42

Howlett NG, Harney JA, Rego MA, Kolling FW, Glover TW

Abstract

Fanconi Anemia (FA) is a rare recessive disease characterized by congenital abnormalities, bone marrow failure, and cancer susceptibility. The FA proteins and the familial breast cancer susceptibility gene products, BRCA1 and FANCD1/BRCA2, function cooperatively in the FA-BRCA pathway to repair damaged DNA and to prevent cellular transformation. Activation of this pathway occurs via the mono-ubiquitination of the FANCD2 protein, targeting it to nuclear foci where it co-localizes with FANCD1/BRCA2, RAD51, and PCNA. The regulation of the mono-ubiquitination of FANCD2, as well as its function in DNA repair remain poorly understood. In this study, we have further characterized the interaction between the FANCD2 and PCNA proteins. We have identified a highly conserved, putative FANCD2 PCNA interaction motif (PIP-box), and demonstrate that mutation of this motif disrupts FANCD2-PCNA binding and precludes the mono-ubiquitination of FANCD2. Consequently, the FANCD2 PIP-box mutant protein fails to correct the mitomycin C hypersensitivity of FA-D2 patient cells. Our results suggest that PCNA may function as a molecular platform to facilitate the mono-ubiquitination of FANCD2 and activation of the FA-BRCA pathway.

MeSH Terms
Animals Apoptosis Regulatory Proteins BRCA2 Protein/metabolism COS Cells Cell Line, Tumor Chlorocebus aethiops DNA Damage DNA Repair Fanconi Anemia/metabolism Fanconi Anemia Complementation Group D2 Protein/metabolism,physiology Humans Mitomycin/chemistry Mutagenesis, Site-Directed Mutation Proliferating Cell Nuclear Antigen/metabolism Ubiquitin/chemistry
Chemicals
Apoptosis Regulatory Proteins BLID protein, human BRCA2 Protein BRCA2 protein, human Fanconi Anemia Complementation Group D2 Protein Proliferating Cell Nuclear Antigen Ubiquitin Mitomycin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Howlett Niall G
Department of Cell and Molecular Biology, University of Rhode Island, Kingston, Rhode Island 02881, USA. [email protected]
Harney Julie A
Rego Meghan A
Kolling Frederick W
Glover Thomas W
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M109.016352
Email
[email protected]
Published
2009-10-16
Epub
2009-00-24
Pages
28935-42
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2781439
Subset
IM
Grants
NCRR NIH HHS · P20 RR016457 · United States
NCI NIH HHS · R01 CA043222 · United States
NCRR NIH HHS · P20RR016457-09 · United States
NCI NIH HHS · R01CA043222-21 · United States
External Links
PubMed source DOI: 10.1074/jbc.M109.016352 PMC Full Text View journal details
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