Characterization of 5'-heterogeneity of the rat GLUT4/muscle-adipose glucose transporter gene product

Ann Louise Olson, Nicholas P. Edgington, W. Scott Moye-Rowley, Jeffrey E. Pessin

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

To examine the mechanisms responsible for tissue-specific, nutritional, and metabolic regulation of the GLUT4/muscle-adipose specific glucose transporter, we isolated and characterized the properties of the rat GLUT4 gene. Examination of the sequenced 2.5-kilobase flanking DNA revealed substantial identity with that of the mouse and human GLUT4 genes, with the greatest degree of sequence identity within the proximal 1000 basepairs up-stream of the GLUT4 open reading frame. Primer extension analysis identified a unique single transcription initiation site 176 basepairs up-stream from the start of translation. However, ribonuclease mapping revealed the presence of a previously undescribed alternatively spliced form of GLUT4 messenger RNA. Approximately 75% of the GLUT4 transcripts consisted of a fully spliced messenger RNA, and 25% was expressed as an unspliced intron-containing species. The ratios of 5' spliced and unspliced messages were invariant in adipose, cardiac, and skeletal muscle tissues. In vitro translation of reporter constructs containing both the spliced and unspliced leader demonstrated a functional difference between these two transcripts, with the unspliced form translated approximately 5-fold more than the fully spliced species. These data demonstrate the presence of 5'-heterogeneity of the GLUT4 transcripts, which underlies differences in translational efficiency in vitro.

Original languageEnglish (US)
Pages (from-to)1962-1968
Number of pages7
JournalEndocrinology
Volume136
Issue number5
DOIs
StatePublished - May 1995
Externally publishedYes

ASJC Scopus subject areas

  • Endocrinology

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