Nucleoside transporter proteins of Saccharomyces cerevisiae. Demonstration of a transporter (FUI1) with high uridine selectivity in plasma membranes and a transporter (FUN26) with broad nucleoside selectivity in intracellular membranes.
FUI1 and function unknown now 26 (FUN26) are proteins of uncertain function with sequence similarities to members of the uracil/allantoin permease and equilibrative nucleoside transporter families of transporter proteins, respectively. [(3)H]Uridine influx was eliminated by disruption of the gene encoding FUI1 (fui1) and restored by expression of FUI1 cDNA, whereas influx in transport-competent and fui1-negative yeast were unaffected, respectively, by disruption of the FUN26 gene or overexpression of FUN26 cDNA. FUI1 transported uridine with high affinity (K(m), 22 +/- 3 micrometer) and was unaffected or inhibited only partially by high concentrations (1 mm) of a variety of ribo- and deoxyribonucleosides or nucleobases. When FUN26 cDNA was expressed in oocytes of Xenopus laevis, inward fluxes of [(3)H]uridine, [(3)H]adenosine, and [(3)H]cytidine were stimulated, and uridine influx was independent of pH and not inhibited by dilazep, dipyridamole, or nitrobenzylmercaptopurine ribonucleoside. Fractionation of yeast membranes containing immunotagged recombinant FUN26 (shown to be functional in oocytes) demonstrated that the protein was primarily in intracellular membranes. These results indicated that FUI1 has high selectivity for uracil-containing ribonucleosides and imports uridine across cell-surface membranes, whereas FUN26 has broad nucleoside selectivity and most likely functions to transport nucleosides across intracellular membranes.
Vickers
M F
MF
Molecular Biology of Membranes Group and Membrane Transport Research Group, Departments of Biochemistry, Oncology, and Physiology, University of Alberta, Canada.
Yao
S Y
SY
Baldwin
S A
SA
Young
J D
JD
Cass
C E
CE
D000818:Animals; D001693:Biological Transport, Active; D005656:Fungal Proteins / Q000378:metabolism*; D007425:Intracellular Membranes / Q000378:metabolism*; D007700:Kinetics; D026901:Membrane Transport Proteins / Q000378:metabolism*; D033703:Nucleoside Transport Proteins; D009865:Oocytes / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D014529:Uridine / Q000378:metabolism*; D014982:Xenopus laevis
C413444:FUI1 protein, S cerevisiae; D005656:Fungal Proteins; D026901:Membrane Transport Proteins; D033703:Nucleoside Transport Proteins; D029701:Saccharomyces cerevisiae Proteins; D014529:Uridine