We intend to define the sequence of human Alpha- and Beta-spectrin mRNAs and characterize the human spectrin genes. Subsequently we shall compare the structure of the spectrin genes from normal individuals and patients with inherited hemolytic anemias in order to identify whether the spectrin gene is altered in such disease. A cloned human Alpha-spectrin cDNA has recently been characterized using a mouse Alpha-spectrin cDNA which had been isolated from a bacterial expression library containing cDNAs from mouse anemic spleen. The same approach will be used to identify a human Beta-spectrin cDNA. From the human cDNAs, full length cDNAs and genomic DNA will be isolated and characterized by restriction site analysis and nucleotide sequencing. With this information, DNA and when possible RNA from patients with inherited hemolytic anemias, particularly those cases where existing evidence implicates spectrin as the site of the defect, will be analyzed in order to identify the genetic basis of these inherited diseases.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL033884-01
Application #
3346202
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1985-05-01
Project End
1990-04-30
Budget Start
1985-05-01
Budget End
1986-04-30
Support Year
1
Fiscal Year
1985
Total Cost
Indirect Cost
Name
Wistar Institute
Department
Type
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
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Ursitti, J A; Kotula, L; DeSilva, T M et al. (1996) Mapping the human erythrocyte beta-spectrin dimer initiation site using recombinant peptides and correlation of its phasing with the alpha-actinin dimer site. J Biol Chem 271:6636-44
Amin, K M; Scarpa, A L; Winkelmann, J C et al. (1993) The exon-intron organization of the human erythroid beta-spectrin gene. Genomics 18:118-25
Kotula, L; DeSilva, T M; Speicher, D W et al. (1993) Functional characterization of recombinant human red cell alpha-spectrin polypeptides containing the tetramer binding site. J Biol Chem 268:14788-93
Kotula, L; Laury-Kleintop, L D; Showe, L et al. (1991) The exon-intron organization of the human erythrocyte alpha-spectrin gene. Genomics 9:131-40
Kotula, L; Curtis, P (1988) Xba I polymorphisms in the 5' region of human erythrocyte alpha spectrin gene (Spna). Nucleic Acids Res 16:10950
Laurila, P; Cioe, L; Kozak, C A et al. (1987) Assignment of mouse beta-spectrin gene to chromosome 12. Somat Cell Mol Genet 13:93-7
Cioe, L; Laurila, P; Meo, P et al. (1987) Cloning and nucleotide sequence of a mouse erythrocyte beta-spectrin cDNA. Blood 70:915-20