The murine 21-hydroxylase (C-21) gene is expressed primarily in the adrenal gland. We propose to further define the features of this gene that direct this tissue-specific expression. Most importantly, however, we propose to identify the regulatory proteins that are expressed in the adrenals that interact with this gene and direct its expression. Specifically, we propose to: 1) Identify the defect in the Y1 cell that prevents production of stable 21-hydroxylase mRNA. 2) Use linker scanning mutagenesis to define the different regions required for 21-hydroxylase-CAT expression in Y1 cells. 3) To test the function of cis-acting regulatory sequences in transgenic mice. 4) Identify derivatives of the Y1 cell line that bear mutations in transacting factor genes. 5) Determine whether transacting factors bind directly to the 21-hydroxylase gene. 6) Clone the genes encoding the transacting factors. Characterization of these proteins and the genes that encode them should provide considerable insight into the process of tissue-specific gene expression.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA046361-03
Application #
3189618
Study Section
Molecular Biology Study Section (MBY)
Project Start
1988-02-01
Project End
1993-01-31
Budget Start
1990-02-01
Budget End
1991-01-31
Support Year
3
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Harvard University
Department
Type
Schools of Medicine
DUNS #
082359691
City
Boston
State
MA
Country
United States
Zip Code
02115
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Cui, Z; Zubiaur, M; Bloch, D B et al. (1991) Expression of a G protein subunit, alpha i-1, in Balb/c 3T3 cells leads to agonist-specific changes in growth regulation. J Biol Chem 266:20276-82
Szyf, M; Schimmer, B P; Seidman, J G (1989) Nucleotide-sequence-specific de novo methylation in a somatic murine cell line. Proc Natl Acad Sci U S A 86:6853-7