In the cerebellum of the mouse the expression of the structural gene (Gdc-1) for the enzyme glycerol-3-phosphate dehydrogenase (GPDH) is modulated by unlinked genes. The most interesting regulatory gene, Gdcr-1, acts by increasing the level of Gdc-1 transcripts. This regulatory gene has two interesting properties. First, it seems to act only in the cerebellum and second, alleles at Gdcr-1 act preferentially on alleles at Gdc-1, i.e., the Gdcr-1c allele preferentially enhances expression of the Gdc-1c allele. In this application we seek to identify sites within or flanking the Gdc-1 gene which is involved in determining the cerebellar specific expression. Available for this project are Gdc-1 genomic clones which were isolated and sequenced in the previous grant period. The experimental approach is to first identify DNA sequences within the Gdc-1 gene or its flanking region which enhance transcription in a transient expression assay. These regions will be evaluated further as the cerebellar regulatory region on the basis of genetic variability and the cerebellar specific expressions in the transient expression assays. A second set of experiments aims to map genetic location of the Gdcr-1 gene and to evaluate whether Gdcr-1 acts in reaggregating cultures of mouse cerebellar cells. A third set of experiments seeks to clone the cDNA and the gene for the embryonic GPDH isozyme which is located on Chromosome 9. The experimental approach is to make a lambdagt 11 cDNA library from rabbit heart and screen for a protein in this library which cross-reacts with anti-mouse embryonic GPDH. The rationale for the experiment is that rabbit heart has isozymes of GPDH which have properties similar to the mouse embryonic enzyme. Once the Gdc-2 gene has been cloned, we will have a chance to identify controlling regions in and around the gene which are involved in expression. A comparison of the controlling regions of Gdc-2 with those associated with Gdc-1 may provide insight into the structural requirement for genes expressed in the embryo as compared to those expressed in the adult.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
2R01HD006712-14
Application #
3310565
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1975-06-01
Project End
1991-05-31
Budget Start
1986-06-01
Budget End
1987-05-31
Support Year
14
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Jackson Laboratory
Department
Type
DUNS #
042140483
City
Bar Harbor
State
ME
Country
United States
Zip Code
04609