Department of Physiology and Biophysics, University of Arkansas for Medical Sciences, Little Rock, Arkansas, USA.
Biophys J. 2012 Feb 22;102(4):768-76. doi: 10.1016/j.bpj.2012.01.005. Epub 2012 Feb 21.
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.
酿酒酵母 SO(4)(=) 转运在很宽的动态范围内受到调节。硫饥饿导致由 Sul1p 和 Sul2p 介导的 (35)SO(4)(=) 内流增加约 10,000 倍;超过 80%的内流是通过 Sul2p 进行的。向 S 饥饿细胞中添加蛋氨酸会导致 SUL1 和 SUL2 mRNA 下降 50 倍(t(1/2)∼5 min),但转运下降较慢(t(1/2)∼1 h)。相比之下,SO(4)(=)的添加不会影响 mRNA,但会导致转运迅速(t(1/2)=2-4 min)下降。在 met3Δ 细胞(无法代谢 SO(4)(=))中,向 S 饥饿细胞中添加 SO(4)(=)会导致 (35)SO(4)(=)内流在细胞 SO(4)(=)含量几乎不变的时间内失活。细胞 SO(4)(=)与转运失活之间的关系表明,细胞 SO(4)(=)不是 Sul2p 失活的信号。相反,转运失活速率与细胞外 SO(4)(=)的依赖性与 (35)SO(4)(=)内流相同,表明 Sul2p 表现出使用依赖性失活;转运过程本身增加了 Sul2p 失活和降解的可能性。此外,在向 S 饥饿的 met3Δ 细胞中添加 >0.02 mM SO(4)(=)后,会短暂地排出 SO(4)(=)。这种短暂的外排提供了对过量 SO(4)(=)内流的进一步保护,可能代表 Sul2p 的另一种转运模式。