Heterologous expression of a mammalian epithelial sodium channel in yeast.
The alpha and beta subunits of the amiloride-sensitive rat epithelial sodium channel (alpha beta ENaC) were expressed in the yeast Saccharomyces cerevisiae. We used a combination of yeast strains, including a mutant in the secretory pathway (sec6), and Western blotting techniques, to show that alpha beta ENaC was synthesized and targeted through the secretory system to the plasma membrane. Yeasts expressing alpha beta ENaC were more sensitive to salt than the parent strain. In addition, amiloride, a specific blocker of ENaC, was found to suppress salt sensitivity in the yeast strain expressing alpha beta ENaC.
Gupta
S S
SS
Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, CT 06510, USA.
Canessa
C M
CM
D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics; D002455:Cell Division / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D003594:Cytoplasmic Granules / Q000187:drug effects / Q000378:metabolism; D053503:Epithelial Sodium Channels; D015966:Gene Expression Regulation, Fungal; D006358:Hot Temperature; D008565:Membrane Proteins; D008826:Microbial Sensitivity Tests; D009154:Mutation / Q000235:genetics; D009994:Osmolar Concentration; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D026941:Sodium Channel Blockers; D015222:Sodium Channels / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012965:Sodium Chloride / Q000037:antagonists & inhibitors / Q000494:pharmacology; D033921:Vesicular Transport Proteins
D002352:Carrier Proteins; D053503:Epithelial Sodium Channels; C096102:Exoc3 protein, rat; D008565:Membrane Proteins; C490684:SEC6 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D026941:Sodium Channel Blockers; D015222:Sodium Channels; D033921:Vesicular Transport Proteins; D012965:Sodium Chloride; D000584:Amiloride