Inactivation of Saccharomyces cerevisiae sulfate transporter Sul2p: use it and lose it

Abstract

Saccharomyces cerevisiae SO(4)(=) transport is regulated over a wide dynamic range. Sulfur starvation causes ∼10,000-fold increase in the (35)SO(4)(=) influx mediated by transporters Sul1p and Sul2p; >80% of the influx is via Sul2p. Adding methionine to S-starved cells causes a 50-fold decline (t(1/2) ∼5 min) in SUL1 and SUL2 mRNA but a slower decline (t(1/2) ∼1 h) in transport. In contrast, SO(4)(=) addition does not affect mRNA but causes a rapid (t(1/2) = 2-4 min) decrease in transport. In met3Δ cells (unable to metabolize SO(4)(=)), addition of SO(4)(=) to S-starved cells causes inactivation of (35)SO(4)(=) influx over times in which cellular SO(4)(=) contents are nearly constant. The relationship between cellular SO(4)(=) and transport inactivation shows that cellular SO(4)(=) is not the signal for Sul2p inactivation. Instead, the transport inactivation rate has the same dependence on extracellular SO(4)(=) as (35)SO(4)(=) influx, indicating that Sul2p exhibits use-dependent inactivation; the transport process itself increases the probability of Sul2p inactivation and degradation. In addition, there is a transient efflux of SO(4)(=) shortly after adding >0.02 mM SO(4)(=) to S-starved met3Δ cells. This transient efflux provides further protection against excessive SO(4)(=) influx and may represent an alternate transport mode of Sul2p.