An evolutionarily conserved N-terminal Sgk1 variant with enhanced stability and improved function

SGK1 is an aldosterone-induced kinase that regulates epithelial Sodium Channel (ENaC)-mediated Na+ transport in the collecting duct and connecting tubule of the kidney. The NH2 terminus of SGK1 contains instability motifs that direct the ubiquitination of SGK1 resulting in a rapidly degraded protein. By bioinformatic analysis, we identified a 5' variant alternate transcript of human SGK1 (Sgk1_v2) that is widely expressed, is conserved from rodent to humans, and is predicted to encode an SGK1 isoform, Sgk1_i2, with a different NH2 terminus. When expressed in HEK293 cells, Sgk1_i2 was more abundant than SGK1 because of an increased protein half-life and this correlated with reduced ubiquitination of Sgk1_i2 and enhanced surface expression of ENaC. Immunocytochemical studies demonstrated that in contrast to SGK1, Sgk1_i2 is preferentially targeted to the plasma membrane. When coexpressed with ENaC subunits in FRT epithelia, Sgk1_i2 had a significantly greater effect on amiloride-sensitive Na+ transport compared with SGK1. Together, the data demonstrate that a conserved NH2-terminal variant of SGK1 shows improved stability, enhanced membrane association, and greater stimulation of epithelial Na+ transport in a heterologous expression system.