The characterization of spontaneous and induced mutants has proven invaluable in understanding mammalian iron metabolism. In particular, positional cloning of genes mutated in mouse, rat, human, and zebrafish iron deficiency and iron overload phenotypes has led to the discovery of multiple proteins that directly participate in or modify iron transport in mammals, many of which are directly relevant to understanding the pathophysiology of human diseases. Over the past several years, the generation of dense genetic and physical maps and near-complete genome sequences of mice and humans has greatly facilitated positional cloning and the rapidity in which mutants can be cloned. Here, we describe the initial phenotypic characterization of a new murine autosomal recessive hypochromic, microcytic anemia mutation, nm1054 (Dew mutation 1054). The phenotype of these animals strongly suggests an underlying defect in erythroid intracellular iron metabolism. Genetic mapping localizes the trait to mouse chromosome 1, and demonstrates that the mutant phenotype is due to a large genomic deletion that includes at least 5 genes. In this grant, we propose to evaluate the hypothesis that nm1054 is a defect in intraerythroid iron metabolism using bone marrow transplantation studies and in vitro iron uptake assays. Furthermore, we will clone the nm1054 anemia gene using bacterial artificial chromosome (BAC) and cDNA transgenic complementation. We expect that determining which of the deleted genes is responsible for the anemia will provide insight into the pathway of iron delivery and utilization in mammalian cells.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL074247-02
Application #
6766851
Study Section
Hematology Subcommittee 2 (HEM)
Program Officer
Evans, Gregory
Project Start
2003-07-01
Project End
2007-06-30
Budget Start
2004-07-01
Budget End
2005-06-30
Support Year
2
Fiscal Year
2004
Total Cost
$405,000
Indirect Cost
Name
Children's Hospital Boston
Department
Type
DUNS #
076593722
City
Boston
State
MA
Country
United States
Zip Code
02115
Lambe, Teresa; Simpson, Robert J; Dawson, Sara et al. (2009) Identification of a Steap3 endosomal targeting motif essential for normal iron metabolism. Blood 113:1805-8
Tian, Meng; Campagna, Dean R; Woodward, Lanette S et al. (2008) hem6: an ENU-induced recessive hypochromic microcytic anemia mutation in the mouse. Blood 112:4308-13
Lee, Lance; Campagna, Dean R; Pinkus, Jack L et al. (2008) Primary ciliary dyskinesia in mice lacking the novel ciliary protein Pcdp1. Mol Cell Biol 28:949-57
Lee, Lance; DeBono, C Anthony; Campagna, Dean R et al. (2007) Loss of the acyl-CoA binding protein (Acbp) results in fatty acid metabolism abnormalities in mouse hair and skin. J Invest Dermatol 127:16-23
Ohgami, Robert S; Campagna, Dean R; McDonald, Alice et al. (2006) The Steap proteins are metalloreductases. Blood 108:1388-94
Ohgami, Robert S; Campagna, Dean R; Antiochos, Brendan et al. (2005) nm1054: a spontaneous, recessive, hypochromic, microcytic anemia mutation in the mouse. Blood 106:3625-31
Ohgami, Robert S; Campagna, Dean R; Greer, Eric L et al. (2005) Identification of a ferrireductase required for efficient transferrin-dependent iron uptake in erythroid cells. Nat Genet 37:1264-9