The Rat Pyruvate Carboxylase Gene Structure ALTERNATE PROMOTERS GENERATE MULTIPLE TRANSCRIPTS WITH THE 5'-END HETEROGNEITY

Simple item page
dc.contributor.authorJitrapakdee, S.
dc.contributor.authorBooker, G.
dc.contributor.authorCassady, A.
dc.contributor.authorWallace, J.
dc.date.issued1997
dc.description.abstractPyruvate carboxylase (EC 6.4.1.1) is a biotin-containing enzyme that plays an important role in gluconeogenesis and lipogenesis. Here we report the structural organization of the rat pyruvate carboxylase gene, which spans over 40 kilobases and is composed of 19 coding exons and 4 5'-untranslated region exons. From this data, it is clear that alternative splicing of the primary transcripts from two promoters is responsible for the occurrence of the multiple mRNA species previously reported (Jitrapakdee, S., Walker, M. E., and Wallace, J. C. (1996) Biochem. Biophys. Res. Commun. 223, 695-700). The proximal promoter, which is active in gluconeogenic and lipogenic tissues, contains no TATA or CAAT boxes but includes a sequence that is typical of a housekeeping initiator protein 1 box while the distal promoter contains three CAAT boxes and multiple Sp1 binding sites. Several potential transcription factor binding sites are found in both promoters. A series of 5'-nested deletion constructs of both promoters were fused to a firefly luciferase reporter plasmid and transiently expressed in COS-1 cells. The results show that the 153 and 187 base pairs, preceding the transcription start sites of the proximal and distal promoters, respectively, are required for basal transcription. Insulin selectively inhibits the expression of the proximal promoter-luciferase reporter gene by 50% but not the distal promoter in COS-1 cells, suggesting the presence of an insulin-responsive element in the proximal promoter. A half-maximal effect was found at approximately 1 nM insulin.
dc.identifier.citationJournal of Biological Chemistry, 1997; 272(33):20522-20530
dc.identifier.doi10.1074/jbc.272.33.20522
dc.identifier.issn0021-9258
dc.identifier.issn1083-351X
dc.identifier.orcidBooker, G. [0000-0001-7207-4699]
dc.identifier.urihttp://hdl.handle.net/2440/11313
dc.language.isoen
dc.publisherAMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
dc.source.urihttps://doi.org/10.1074/jbc.272.33.20522
dc.subjectAnimals
dc.subjectRats
dc.subjectInsulin
dc.subjectPyruvate Carboxylase
dc.subjectRNA, Messenger
dc.subjectPolymerase Chain Reaction
dc.subjectAlternative Splicing
dc.subjectBase Sequence
dc.subjectExons
dc.subjectMolecular Sequence Data
dc.subjectPromoter Regions, Genetic
dc.titleThe Rat Pyruvate Carboxylase Gene Structure ALTERNATE PROMOTERS GENERATE MULTIPLE TRANSCRIPTS WITH THE 5'-END HETEROGNEITY
dc.typeJournal article
pubs.publication-statusPublished

Files

Collections

Biochemistry publications