CRIS NUMBER: 0180820
SUBFILE: CRIS
PROJECT NUMBER: CA-V*-PHR-6562-H
SPONSOR AGENCY: CSREES
PROJECT TYPE: HATCH
PROJECT STATUS: TERMINATED
MULTI-STATE PROJECT NUMBER: (N/A)
START DATE: Jan 1, 1999
TERMINATION DATE: Dec 31, 2003
GRANT PROGRAM: (N/A)
GRANT PROGRAM AREA: (N/A)
CLASSIFICATION
311 | 3510 | 1080 | 4.2 | 25% |
311 | 3510 | 1090 | 4.2 | 25% |
311 | 3510 | 1102 | 4.2 | 25% |
311 | 3510 | 1020 | 4.2 | 25% |
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CLASSIFICATION HEADINGS
KA311 - Animal Diseases S3510 - Swine, live animal F1020 - Physiology F1080 - Genetics F1102 - Mycology F1090 - Immunology G4.2 - Reduce Number and Severity of Pest and Disease Outbreaks
RESEARCH EFFORT CATEGORIES
BASIC |
(N/A)% |
APPLIED |
100% |
DEVELOPMENTAL |
(N/A)% |
KEYWORDS: differentiation; promoters (genetics); genetic regulation; animal genetics; in situ hybridization; swine; pregnancy; transcriptional regulation; transcription; gene expression; binding proteins; blastocysts; dna sequences; reporter genes; protein binding; protein purification; protein characterization; mutagenesis; gene analysis
PROGRESS: Jan 1, 1999 TO Dec 31, 2003
Trophoblastic differentiation is essential for the establishment and successful completion of pregnancy. This process proceeds in porcine pre-attachment blastocysts with the expression of the CYP17 gene that encodes for the enzyme 17 -hydroxylase/17,20-lyase cytochrome P450 (P450c17). The expression of this enzyme, exclusively in the trophectoderm layer, is associated with trophoblastic differentiation and is essential for conceptus survival in utero. We have cloned the entire CYP17 gene in the domestic pig, and have begun defining sequence elements in the regulatory region that are unique in driving trophoblastic expression. The specific aims of the project are: 1) To define the core sequence of the palindromic promoter driving CYP17 expression in porcine trophoblast, 2) To clone transcription activating protein(s) associated with the core palindromic promoter. Results We have defined a novel, non-TATA promoter element directing the assembly of a transcriptional
complex at a position on the gene that is uniquely used for the expression of CYP17 in the porcine trophectoderm. The essential core element exists within a 34 base pair fragment of the porcine CYP17 gene, 142 bp 5' of the transcription start site utilized in gonadal and adrenal tissues. It has extensive homology to previously described 'initiator elements', but with a very different core structure. Site-directed mutagenesis experiments, together with gel shift and southwestern blotting experiments have defined the essential nucleotide bases associated with promoter activity. These do not correspond to those identified as core for initiator elements. Finally, we have only just begun screening of expression libraries, with a plan to use our defined promoter element, and inactive point mutants, as probes to clone the protein(s) that bind and thereby modulate the activity of his unique trophoblastic porcine CYP17 promoter. These experiments will completed, and the final goal of the
project pursued further, even though the current project will be officially terminated with this report.
IMPACT: 1999-01-01 TO 2003-12-31
The placenta is the organ that determines the growth rate and well-being of the fetus, and diseases or conditions such as the large calf syndrome associated with cloned animals are believed to arise from derangements of placental development. The research described adds to our understanding of the transcriptional and other regulatory mechanisms that are unique to the placenta and therefore are likely to provide insight into diseases afflicting offspring before and after birth.
PUBLICATION INFORMATION: 1999-01-01 TO 2003-12-31
No publications reported this period
PROJECT CONTACT INFORMATION
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