The canine sodium/myo-inositol cotransporter gene: structural organization and characterization of the promoter.
The sodium/myo-inositol cotransporter (SMIT) is a plasma membrane protein catalyzing transfer of myo-inositol into cells against a considerable concentration gradient using the electrochemical potential of sodium across the cell membrane. Transcription of the SMIT gene is markedly stimulated when cells are exposed to a hypertonic environment resulting in increased abundance of SMIT mRNA and increased SMIT activity. The increased accumulation of myo-inositol protects cells from the deleterious effects of hypertonicity. In an effort toward understanding transcriptional regulation, we cloned canine genomic DNA fragments containing the SMIT gene. The gene is 37 kb in size consisting of 2 exons and a large intron of 25 kb. The entire open reading frame is in the second exon. The promoter of the gene is highly active due to a GC-rich sequence. Ribonuclease protection assay using a riboprobe complementary to the 5' end of the gene confirmed that the promoter of the gene is stimulated by hypertonicity. The promoters and regulatory sequences of the SMIT gene and the betaine transporter gene, another gene regulated by hypertonicity, appear to be different.
Rim
J S
JS
Division of Nephrology, Johns Hopkins School of Medicine, Baltimore, Maryland 21205, USA.
Tanawattanacharoen
S
S
Takenaka
M
M
Handler
J S
JS
Kwon
H M
HM
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000235:genetics*; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004285:Dogs; D005091:Exons / Q000235:genetics; D015723:Gene Library; D017930:Genes, Reporter; D006360:Heat-Shock Proteins / Q000235:genetics*; D007294:Inositol / Q000378:metabolism; D007438:Introns / Q000235:genetics; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D027981:Symporters; D014158:Transcription, Genetic / Q000235:genetics
D002352:Carrier Proteins; D006360:Heat-Shock Proteins; D008565:Membrane Proteins; D027981:Symporters; C073884:SLC5A3 protein, human; D007294:Inositol; D008156:Luciferases; D015500:Chloramphenicol O-Acetyltransferase