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J Biol Chem, Vol. 274, Issue 26, 18741-18746, June 25, 1999
From the Bakers' yeast is auxotrophic for biotin (vitamin
H) and depends on the efficient uptake of this compound from the
environment. A mutant strain with strongly reduced biotin uptake and
with reduced levels of protein biotinylation was identified. The strain
was auxotrophic for long-chain fatty acids, and this auxotrophy could be suppressed with high levels of biotin in the medium. After transformation of this mutant with a yeast genomic library, the unassigned open reading frame YGR065C was identified to
complement this mutation. This gene codes for a protein with 593 amino
acids and 12 putative transmembrane helices. Northern blot analysis revealed that, in wild-type cells, the corresponding mRNA levels were increased at low biotin concentrations. Likewise, cellular biotin
uptake was increased with decreasing biotin availability. Expression of
YGR065C under the control of the constitutive
ADH1 promoter resulted in very high biotin transport rates
across the plasma membrane that were no longer regulated by the biotin
concentration in the growth medium. We conclude that
YGR065C encodes the first biotin transporter identified for
a non-mammalian organism and designate this gene VHT1 for
vitamin H transporter
1.
Identification of the Plasma Membrane H+-Biotin
Symporter of Saccharomyces cerevisiae by Rescue of a Fatty
Acid-auxotrophic Mutant
,, and Lehrstuhl Botanik II,
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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