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dc.contributor.authorJung, Yong-keun
dc.identifier.citationSource: Dissertation Abstracts International, Volume: 54-04, Section: B, page: 1797.;Advisors: Lloyd D. Fricker.
dc.description.abstractThe gene for rat carboxypeptidase E (CPE), a neuropeptide-processing enzyme, has been cloned and partially sequenced. The entire CPE gene spans 50 kilobases and consist of nine exons, each of which contains protein-coding regions. Only one exon/intron junction of the rat CPE gene is present in a comparable position within the genes for carboxypeptidase-A and -B. Southern blot analysis indicates that a single copy of this gene is present in the rat genome. Nuclease protection shows that no alternative splicing of exons 7, 8, and 9 occurs, indicating that heterogeneity previously found within the C-terminal region of CPE is due to posttranslational processing.;Analyses have identified a single major transcription start site for CPE mRNA in most tissues. The promoter of CPE does not contain TATA and CAAT boxes but has an initiator-like sequence around the major transcription start site. Additional pituitary-specific transcription start sites {dollar}(-{dollar}101 and {dollar}-{dollar}105) were identified.;The CPE promoter (up to {dollar}-{dollar}12kb) has been analyzed to identify cis-acting elements involved with the expression of the rat CPE gene. A portion of the CPE gene spanning the major transcription start site {dollar}(-{dollar}12 to +47) is sufficient for low levels of promoter activity. Reporter gene activities are enhanced 3-15 fold by the sequences between {dollar}-{dollar}12 and {dollar}-{dollar}395 in all cell lines and the activities were influenced up to 5-fold by the sequences between {dollar}-{dollar}395 and {dollar}-{dollar}3081 in some cell lines. There is no correlation between transcription activity of the various constructs and the level of endogenous CPE mRNA in the cell lines, indicating that the major tissue-specific elements responsible for the large variation in endogenous CPE mRNA are not present within {dollar}-{dollar}12,000 to +47.;DNase protection analysis using {dollar}\rm GH\sb4C\sb1{dollar} nuclear extract shows many DNA binding factors which recognize consensus sequences and transcription start sites in the {dollar}-{dollar}350 to +80 region. The CPE promoter region spanning {dollar}-{dollar}12 to +20 is a functional initiator, showing an orientation-dependent expression pattern. A distinct CPE initiator-binding protein was detected, indicating that CPE initiator element is recognized by a novel initiator-binding factor and more than a single initiator-binding protein is present in {dollar}\rm GH\sb4C\sb1{dollar} cells. (Abstract shortened by UMI.).
dc.publisherProQuest Dissertations & Theses
dc.subjectMolecular biology.
dc.titleRat carboxypeptidase E gene: Structure, expression, and regulation of expression

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