The study of rare genetic disorders of cancer susceptibility, such as Fanconi anemia (FA) have led to seminal advances in cancer biology as a whole. The hallmark of FA is genomic instability and DNA damage hypersensitivity, but the normal function of the FA pathway or mechanism leading to cancer in the affected patients remains unknown. FA is accounted for by multiple complementation groups corresponding to at least 8 cloned genes whose encoded proteins contain few functional motifs. What is known is that the FA pathway interacts with other more defined pathways involved in cellular response to DNA damage, such as ATM and BRCA1. The Kupfer laboratory has established that FA proteins such as FANCA and FANCG phosphorylated, that the FA core complex (composed of at least 5 FA proteins) localizes to at least 3 subcellular compartments, including chromatin, and that the core complex binds to additional non-characterized proteins. This proposal centers on the analysis of posttranslational modifications of the FA proteins FANCA and FANCG and on detecting new binding proteins and determining their function. This would shed light on the long-term goal of the Kupfer laboratory: define the mechanisms of genomic instability caused by FA as well as understand the normal function of the FA proteins. In this proposal the experiments proposed address the central hypothesis: that the phosphorylation of FANCA and FANCG regulate proper localization to chromatin and that a novel FA binding protein, RBM10, has RNA binding activity critical for this localization. The work proposed will 1) determine the cause and effect of FANCA and FANCG phosphorylation, 2) analyze RBM10 and RNA function in the FA core complex, and 3) focus on purifying additional FA core complex and FANCD2 binding proteins, especially from chromatin. This research is significant because it sheds light on a mechanism of genomic instability at the level of the functional genetic unit of the cell: chromatin. Because the response to DNA damage involves numerous intersecting pathways, defining the function of FA proteins will allow us to link these disparate pathways in a mechanism explaining cancer formation.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
7R01HL063776-08
Application #
7591461
Study Section
Hematopoiesis Study Section (HP)
Program Officer
Qasba, Pankaj
Project Start
2000-01-01
Project End
2010-05-31
Budget Start
2007-09-05
Budget End
2008-05-31
Support Year
8
Fiscal Year
2007
Total Cost
$273,969
Indirect Cost
Name
Yale University
Department
Pediatrics
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Longerich, Simonne; Kwon, Youngho; Tsai, Miaw-Sheue et al. (2014) Regulation of FANCD2 and FANCI monoubiquitination by their interaction and by DNA. Nucleic Acids Res 42:5657-70
Chen, Xiaoyong; Wilson, James B; McChesney, Patricia et al. (2014) The Fanconi anemia proteins FANCD2 and FANCJ interact and regulate each other's chromatin localization. J Biol Chem 289:25774-82
Chirnomas, S Deborah; Kupfer, Gary M (2013) The inherited bone marrow failure syndromes. Pediatr Clin North Am 60:1291-310
Kupfer, Gary M (2013) Fanconi anemia: a signal transduction and DNA repair pathway. Yale J Biol Med 86:491-7
Williams, Stacy A; Longerich, Simonne; Sung, Patrick et al. (2011) The E3 ubiquitin ligase RAD18 regulates ubiquitylation and chromatin loading of FANCD2 and FANCI. Blood 117:5078-87
Williams, Stacy A; Wilson, James B; Clark, Allison P et al. (2011) Functional and physical interaction between the mismatch repair and FA-BRCA pathways. Hum Mol Genet 20:4395-410
Zhi, Gang; Chen, Xiaoyong; Newcomb, William et al. (2010) Purification of FANCD2 sub-complexes. Br J Haematol 150:88-92
Wilson, James B; Blom, Eric; Cunningham, Ryan et al. (2010) Several tetratricopeptide repeat (TPR) motifs of FANCG are required for assembly of the BRCA2/D1-D2-G-X3 complex, FANCD2 monoubiquitylation and phleomycin resistance. Mutat Res 689:12-20
Song, Ihn Young; Palle, Komaraiah; Gurkar, Aditi et al. (2010) Rad18-mediated translesion synthesis of bulky DNA adducts is coupled to activation of the Fanconi anemia DNA repair pathway. J Biol Chem 285:31525-36
Zhi, Gang; Wilson, James B; Chen, Xiaoyong et al. (2009) Fanconi anemia complementation group FANCD2 protein serine 331 phosphorylation is important for fanconi anemia pathway function and BRCA2 interaction. Cancer Res 69:8775-83

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