Linked Life Data

9490043

Source:http://linkedlifedata.com/resource/pubmed/id/9490043

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
3
pubmed:dateCreated
1998-3-23
pubmed:abstractText
Transgenic mice were previously described, carrying the cDNA of the human or murine protease urokinase-type plasminogen activator (uPA) while linked to the cell-specific promoter of the alphaA-crystallin gene. Surprisingly, these mice produced transgenic uPA in an ectopic manner specifically in the brain. Here we tested the possibility that this ectopic expression could have been contributed primarily by the uPA transgenic moiety. Several experimental approaches have been used. (a) Constructs consisting of uPA cDNA linked to the cell-specific promoters of the alphaA-crystallin or insulin genes yielded active uPA after transfection into cells where these promoters are thought to be inactive. (b) When reporter genes were inserted into these constructs between the promoter and the cDNA, the cDNA enhanced the chimeric reporter expression 5-50-fold. This effect was obtained upon stable or transient construct transfection into four different cell types. (c) Reporter enhancement also took place in the presence of the homologous uPA gene promoter. (d) Mapping of the cDNA through deletion-substitution analysis has detected fragments mediating positive or negative effects on reporter expression, all fragments residing in the 3'-untranslated region (3'UTR) of the uPA gene that was included in the cDNA. Some fragments exhibited cell-specific effects. One fragment (2002/2187) behaved like a classical transcriptional enhancer, enhancing reporter expression from different positions and orientations. (e) Transgenic mice have now been generated that carry a transgene consisting of the alphaA-crystallin promoter, the luciferase reporter gene and mouse uPA cDNA. Among four transgenic lines producing luciferase activity in the eye lens, three lines exhibited ectopic luciferase activity exclusively in the brain, where luciferase mRNA was localized through in situ hybridization. From these results we conclude that the 3'UTR of the uPA gene contains sequences capable of exerting variable effects on gene expression, including transcriptional enhancement. In addition, uPA cDNA correlates with transgenic brain expression. Therefore, we suggest that the 3'UTR of the uPA gene is involved in brain expression of the transgenes containing uPA cDNA as well as of the normal uPA gene.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0014-2956
pubmed:author
pubmed:issnType
Print
pubmed:day
1
pubmed:volume
251
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
704-15
pubmed:dateRevised
2011-11-17
pubmed:meshHeading
pubmed-meshheading:9490043-Animals, pubmed-meshheading:9490043-Brain, pubmed-meshheading:9490043-Chloramphenicol O-Acetyltransferase, pubmed-meshheading:9490043-Crystallins, pubmed-meshheading:9490043-DNA, Complementary, pubmed-meshheading:9490043-Electroporation, pubmed-meshheading:9490043-Humans, pubmed-meshheading:9490043-Insulin, pubmed-meshheading:9490043-Luciferases, pubmed-meshheading:9490043-Mice, pubmed-meshheading:9490043-Mice, Transgenic, pubmed-meshheading:9490043-Organ Specificity, pubmed-meshheading:9490043-Promoter Regions, Genetic, pubmed-meshheading:9490043-Recombinant Fusion Proteins, pubmed-meshheading:9490043-Restriction Mapping, pubmed-meshheading:9490043-Sequence Deletion, pubmed-meshheading:9490043-Transfection, pubmed-meshheading:9490043-Urokinase-Type Plasminogen Activator
pubmed:year
1998
pubmed:articleTitle
The 3'-untranslated region of the urokinase gene enhances the expression of chimeric genes in cultured cells and correlates with specific brain expression in transgenic mice.
pubmed:affiliation
Department of Biochemistry, The Weizmann Institute of Science, Rehovot, Israel.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't