Tpn1p, the plasma membrane vitamin B6 transporter of Saccharomyces cerevisiae.
Pyridoxine (PN) is a metabolic precursor of pyridoxal phosphate that functions as a cofactor of many enzymes in amino acid metabolism. PN, pyridoxal, and pyridoxamine are collectively referred to as vitamin B6, and mammalian organisms depend on its uptake from the diet. In addition to the ability to use extracellular vitamin B6, most unicellular organisms are also capable of synthesizing PN to generate pyridoxal phosphate. Here, we report the isolation of Saccharomyces cerevisiae mutants that have lost the ability to transport PN across the plasma membrane. We used these mutants to isolate TPN1, the first known gene encoding a transport protein for vitamin B6. Tpn1p is a member of the purine-cytosine permease family within the major facilitator superfamily. The protein functions as a proton symporter, localizes to the plasma membrane, and has high affinity for PN. TPN1 mutants lost the ability to utilize extracellular PN, pyridoxal, and pyridoxamine, showing that there is no other transporter for vitamin B6 encoded in the genome. Amino acid substitutions that led to a loss of Tpn1p function localized to transmembrane domain 4 within the 12-transmembrane domain protein. Moreover, expression of TPN1 was regulated and increased with decreasing concentrations of vitamin B6 in the medium. We also provide evidence that of the highly conserved SNZ and SNO genes in S. cerevisiae, only the protein encoded by SNZ1 is required for vitamin B6 biosynthesis.
Stolz
Jürgen
J
Lehrstuhl für Zellbiologie und Pflanzenphysiologie, Universität Regensburg, Universitätsstrasse 31, D-93040 Regensburg, Germany. juergen.stolz@biologie.uni-regensburg.de
Vielreicher
Martin
M
D000483:Alleles; D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000378:metabolism; D003433:Crosses, Genetic; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic; D011730:Pyridoxal / Q000378:metabolism; D011732:Pyridoxal Phosphate / Q000378:metabolism; D011733:Pyridoxamine / Q000378:metabolism; D011736:Pyridoxine / Q000008:administration & dosage / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013329:Structure-Activity Relationship; D014316:Tritium; D025101:Vitamin B 6 / Q000378:metabolism*
D008164:Luminescent Proteins; D026901:Membrane Transport Proteins; D029701:Saccharomyces cerevisiae Proteins; C475166:TPN1 protein, S cerevisiae; D014316:Tritium; D049452:Green Fluorescent Proteins; D011730:Pyridoxal; D011732:Pyridoxal Phosphate; D011733:Pyridoxamine; D025101:Vitamin B 6; D011736:Pyridoxine