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J Biol Chem, Vol. 273, Issue 5, 2829-2834, January 30, 1998
From the Department of Biochemistry and the Lucille P. Markey
Cancer Center, University of Kentucky Medical Center,
Lexington, Kentucky 40536-0084
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.
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