Cloning of a Na(+)- and Cl(-)-dependent betaine transporter that is regulated by hypertonicity.
Many hypertonic bacteria, plants, marine animals, and the mammalian renal medulla are protected from the deleterious effects of high intracellular concentrations of electrolytes by accumulating high concentrations of the nonperturbing osmolyte betaine. When kidney-derived Madin-Darby canine kidney (MDCK) cells are cultured in hypertonic medium, they accumulate betaine to 1,000 times its medium concentration. This results from induction by hypertonicity of high rates of betaine transport into cells. We have isolated a cDNA (BGT-1) encoding a renal betaine transporter by screening an MDCK cell cDNA library for expression of a betaine transporter in Xenopus oocytes. The cDNA encodes a single protein of 614 amino acids, with an estimated molecular weight of 69 kDa. The deduced amino acid sequence exhibits highly significant sequence and topographic similarity to brain gamma-amino-n-butyric acid (GABA) and noradrenaline transporters, suggesting that the renal BGT-1 is a member of the brain GABA/noradrenaline transporter gene family. Expression in oocytes indicates that the BGT-1 protein has both betaine and GABA transport activities that are Cl(-)- as well as Na(+)-dependent and functionally similar to betaine and GABA transport in MDCK cells. Northern hybridization indicates that transporter mRNA is localized to the kidney medulla and is induced in MDCK cells by hypertonicity.
Yamauchi
A
A
Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Uchida
S
S
Kwon
H M
HM
Preston
A S
AS
Robey
R B
RB
Garcia-Perez
A
A
Burg
M B
MB
Handler
J S
JS
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001622:Betaine / Q000378:metabolism*; D001692:Biological Transport; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000235:genetics*; D002478:Cells, Cultured; D002712:Chlorides / Q000378:metabolism*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004285:Dogs; D050485:GABA Plasma Membrane Transport Proteins; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D008969:Molecular Sequence Data; D009638:Norepinephrine / Q000378:metabolism; D009994:Osmolar Concentration; D010957:Plasmids; D011061:Poly A / Q000378:metabolism; D011487:Protein Conformation; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012964:Sodium / Q000378:metabolism*; D014981:Xenopus; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
D002352:Carrier Proteins; D002712:Chlorides; D050485:GABA Plasma Membrane Transport Proteins; D012333:RNA, Messenger; C072207:betaine plasma membrane transport proteins; D011061:Poly A; D001622:Betaine; D005680:gamma-Aminobutyric Acid; D012313:RNA; D004247:DNA; D012964:Sodium; D009638:Norepinephrine