Inhibition of amino acid transport by sphingoid long chain bases in Saccharomyces cerevisiae.
Sphingoid long chain bases have many effects on cells including inhibition or stimulation of growth. The physiological significance of these effects is unknown in most cases. To begin to understand how these compounds inhibit growth, we have studied Saccharomyces cerevisiae cells. Growth of tryptophan (Trp-) auxotrophs was more strongly inhibited by phytosphingosine (PHS) than was growth of Trp+ strains, suggesting that PHS diminishes tryptophan uptake and starves cells for this amino acid. This hypothesis is supported by data showing that growth inhibition is relieved by increasing concentrations of tryptophan in the culture medium and by multiple copies of the TAT2 gene, encoding a high affinity tryptophan transporter. Measurement of tryptophan uptake shows that it is inhibited by PHS. Finally, PHS treatment induces the general control response, indicating starvation for amino acids. Multiple copies of TAT2 do not protect cells against two other cationic lipids, stearylamine, and sphingosine, indicating that the effect of PHS on tryptophan utilization is specific. Other data demonstrate that PHS reduces uptake of leucine, histidine, and proline by specific transporters. Our data suggest that PHS targets proteins in the amino acid transporter family but not other distantly related membrane transporters, including those necessary for uptake of adenine and uracil.
Skrzypek
M S
MS
Department of Biochemistry and the Lucille P. Markey Cancer Center, University of Kentucky Medical Center, Lexington, Kentucky 40536-0084, USA.
Nagiec
M M
MM
Lester
R L
RL
Dickson
R C
RC
D026905:Amino Acid Transport Systems; D000596:Amino Acids / Q000378:metabolism*; D000598:Amino Acids, Cyclic / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects; D002352:Carrier Proteins / Q000187:drug effects* / Q000378:metabolism; D029968:Escherichia coli Proteins; D007930:Leucine / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000187:drug effects* / Q000378:metabolism; D011392:Proline / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000494:pharmacology
D026905:Amino Acid Transport Systems; D000596:Amino Acids; D000598:Amino Acids, Cyclic; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; C035131:TnaB protein, E coli; D011392:Proline; C012491:phytosphingosine; D007930:Leucine; D013110:Sphingosine