The pub1 E3 ubiquitin ligase negatively regulates leucine uptake in response to NH(4)(+) in fission yeast.
Fission yeast strains auxotrophic for leucine are unable to proliferate in normally supplemented minimal media adjusted to pH 6. 4 or above. High-pH sensitivity can be suppressed by the loss of Pub1, an E3 ubiquitin ligase, or by the replacement of NH(4)(+) with a non-repressing source of nitrogen such as L-proline. In this report we show pub1 to be required for the rapid down-regulation of leucine uptake observed in response to the addition of NH(4)(+) to the growth media. Furthermore, we corroborate earlier results demonstrating the transport of leucine to be negatively influenced by high extracellular pH. pub1 is homologous to the budding yeast nitrogen permease inactivator, NPI1/RSP5, which mediates the ubiquitination and subsequent destruction of NH(4)(+)-sensitive permeases. The high-pH sensitivity of cells auxotrophic for leucine thus seems to reflect an inability of NH(4)(+)-insensitive permeases to transport sufficient leucine under conditions where the proton gradient driving nutrient transport is low, and NH(4)(+)-sensitive permeases have been destroyed. Intriguingly, the partial suppression of both high pH sensitivity, and the inactivating effect of NH(4)(+) on leucine transport, seen in pub1-1 point mutants, becomes as complete as seen in pub1Delta backgrounds when cells have concomitantly lost the function of the spc1 stress-activated MAPK.
Karagiannis
J
J
Department of Biology, Queen's University, Kingston, Ontario, K7L 3N6 Canada.
Saleki
R
R
Young
P G
PG
D002455:Cell Division / Q000187:drug effects; D003470:Culture Media / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007930:Leucine / Q000235:genetics / Q000493:pharmacokinetics* / Q000494:pharmacology; D008025:Ligases / Q000235:genetics / Q000502:physiology*; D009154:Mutation; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology*; D012568:Schizosaccharomyces / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases; D014498:Uracil / Q000494:pharmacology
D003470:Culture Media; D000644:Quaternary Ammonium Compounds; D014498:Uracil; D044767:Ubiquitin-Protein Ligases; D008025:Ligases; D007930:Leucine