The objective of this proposal is to understand the role of frascati, a novel mitochondrial solute transporter (SLC25), in vertebrate erythropoiesis and iron metabolism. Acquired and congential defects in iron metabolism from either deficiency or excess are one of the most common medical diseases. Mutations in the zebrafish frascati gene lead to a profound hypochromic anemia and a developmental arrest at the pro- erythroblast stage. Using a positional cloning strategy, we showed that frascati encodes a member of the mitochondrial solute carrier family. The frascati gene (mitoferrin, slc25a37) is highly expressed in fetal and adult hematopoietic tissues of zebrafish and mouse. We hypothesize that frascati is responsible for delivering iron to the mitochondria, and in particular, the mitochondria of developing erythroid cells. The goal of this application is to test this hypothesis. We propose to accomplish this goal by the following two specific aims:
Specific Aim 1 : Analysis of the biochemical function of the frascati gene. 1.1 The role of the frascati gene in mitochondrial iron acquisition will be studied using immortalized cells and zebrafish embryos deficient in frascati activity. 1.2 The physiologic substrate and transport properties of the frascati mitochondrial transporter will be studied by expression of recombinant protein as inclusion bodies in E. coli and their reconstitution in proteoliposomes.
Specific Aim 2 : Analysis of the frascati gene in mammalian organisms. 2.1 The function of the frascati gene in mouse will be studied by targeted gene disruption. Insight into the function of the frascati gene will be directly relevant to our understanding of human disorders of iron deficiency anemia and iron-overload.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK070838-04
Application #
7802269
Study Section
Erythrocyte and Leukocyte Biology Study Section (ELB)
Program Officer
Bishop, Terry Rogers
Project Start
2007-02-01
Project End
2012-01-31
Budget Start
2010-02-01
Budget End
2011-01-31
Support Year
4
Fiscal Year
2010
Total Cost
$348,059
Indirect Cost
Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115
Chung, Jacky; Wittig, Johannes G; Ghamari, Alireza et al. (2017) Erythropoietin signaling regulates heme biosynthesis. Elife 6:
Kafina, Martin D; Paw, Barry H (2017) Intracellular iron and heme trafficking and metabolism in developing erythroblasts. Metallomics 9:1193-1203
Seguin, Alexandra; Takahashi-Makise, Naoko; Yien, Yvette Y et al. (2017) Reductions in the mitochondrial ABC transporter Abcb10 affect the transcriptional profile of heme biosynthesis genes. J Biol Chem 292:16284-16299
Grillo, Anthony S; SantaMaria, Anna M; Kafina, Martin D et al. (2017) Restored iron transport by a small molecule promotes absorption and hemoglobinization in animals. Science 356:608-616
Yien, Yvette Y; Ducamp, Sarah; van der Vorm, Lisa N et al. (2017) Mutation in human CLPX elevates levels of ?-aminolevulinate synthase and protoporphyrin IX to promote erythropoietic protoporphyria. Proc Natl Acad Sci U S A 114:E8045-E8052
van der Vorm, Lisa N; Paw, Barry H (2017) Studying disorders of vertebrate iron and heme metabolism using zebrafish. Methods Cell Biol 138:193-220
Lichtenstein, Daniel A; Crispin, Andrew W; Sendamarai, Anoop K et al. (2016) A recurring mutation in the respiratory complex 1 protein NDUFB11 is responsible for a novel form of X-linked sideroblastic anemia. Blood 128:1913-1917
Yien, Yvette Y; Paw, Barry H (2016) A role for iron deficiency in dopaminergic neurodegeneration. Proc Natl Acad Sci U S A 113:3417-8
Chung, Jacky; Bauer, Daniel E; Ghamari, Alireza et al. (2015) The mTORC1/4E-BP pathway coordinates hemoglobin production with L-leucine availability. Sci Signal 8:ra34
Shmukler, Boris E; Huston, Nicholas C; Thon, Jonathan N et al. (2015) Homozygous knockout of the piezo1 gene in the zebrafish is not associated with anemia. Haematologica 100:e483-5

Showing the most recent 10 out of 48 publications