We plan to investigate five aspects of the carbonic anhydrase (CA) multigene family in humans and other organisms: (1) the characterization of the structure, organization, and evolution of the CA gene family, (2) definition of DNA sequences important in the tissue specific and developmental regulation of mammalian CA genes, (3) examination of the molecular bases of the mutations responsible for inherited deficiencies of CA isozymes, (4) investigation of the role of active site residues in the activities of CA isozymes with different properties, and (5) structural and functional analysis of a CA-like vaccinia virus gene and its product. In the first area, we will complete the characterization of the CA genes we have already cloned (human CA II, macaque CA I, human CA Z, and two different but as yet unidentified human genes) in addition to screening for and characterizing new CA genes. Regulatory sequences in CA genes will be investigated by comparative sequence analysis, construction of deletion mutants, transfection studies and transgenic mice experiments to elucidate those sequences which direct CA gene expression in healthy, functioning organisms. Conversely, in the third area we will study the molecular lesions which cause deficiencies in expression of the CA isozymes such as the human CA II deficiency syndrome, CA II deficient mouse model and the macaque CA I deficiency. The human CA II deficiency results in a syndrome characterized by dysfunction of bone (osteopetrosis), kidney (renal tubular acidosis), and brain (cerebral calcification) and suggest that deficiencies in other CA isozymes may be causative factors in undefined genetic diseases. In the fourth area, we will construct mutations by site-directed mutagenesis in the codons for residues believed to be important in the catalytic mechanisms of different CA isozymes, express them in E. coli, and carry out kinetic studies on the purified altered enzymes. Finally, we will study a vaccinia virus gene and its product, the derived sequence of which is highly homologous to the CAs, to gain insights into its origin, function, and evolutionary relationship to CA genes in other organisms.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM024681-12
Application #
3272443
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1977-12-01
Project End
1992-12-31
Budget Start
1989-01-01
Budget End
1989-12-31
Support Year
12
Fiscal Year
1989
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Type
Schools of Medicine
DUNS #
791277940
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Porter, Calvin A; Hewett-Emmett, David; Tashian, Richard E (2013) Carbonic anhydrase-related protein XI: structure of the gene in the greater false vampire bat (Megaderma lyra) compared with human and domestic pig. Biochem Genet 51:474-81
Yang, H; Hewett-Emmett, D; Tashian, R E (2000) Absence or reduction of carbonic anhydrase II in the red cells of the beluga whale and llama: implications for adaptation to hypoxia. Biochem Genet 38:241-52
Tashian, R E; Hewett-Emmett, D; Carter, N et al. (2000) Carbonic anhydrase (CA)-related proteins (CA-RPs), and transmembrane proteins with CA or CA-RP domains. EXS :105-20
Earnhardt, J N; Qian, M; Tu, C et al. (1998) The catalytic properties of murine carbonic anhydrase VII. Biochemistry 37:10837-45
Yang, H; Nevo, E; Tashian, R E (1998) Unexpected expression of carbonic anhydrase I and selenium-binding protein as the only major non-heme proteins in erythrocytes of the subterranean mole rat (Spalax ehrenbergi). FEBS Lett 430:343-7
Lovejoy, D A; Hewett-Emmett, D; Porter, C A et al. (1998) Evolutionarily conserved, ""acatalytic"" carbonic anhydrase-related protein XI contains a sequence motif present in the neuropeptide sauvagine: the human CA-RP XI gene (CA11) is embedded between the secretor gene cluster and the DBP gene at 19q13.3. Genomics 54:484-93
Lakkis, M M; Venta, P J; Tashian, R E (1997) Localization of the mitochondrial carbonic anhydrase V gene, Car5, on mouse chromosome 8. Mamm Genome 8:225-6
Lakkis, M M; Bergenhem, N C; O'Shea, K S et al. (1997) Expression of the acatalytic carbonic anhydrase VIII gene, Car8, during mouse embryonic development. Histochem J 29:135-41
Lakkis, M M; O'Shea, K S; Tashian, R E (1997) Differential expression of the carbonic anhydrase genes for CA VII (Car7) and CA-RP VIII (Car8) in mouse brain. J Histochem Cytochem 45:657-62
Hewett-Emmett, D; Tashian, R E (1996) Functional diversity, conservation, and convergence in the evolution of the alpha-, beta-, and gamma-carbonic anhydrase gene families. Mol Phylogenet Evol 5:50-77

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