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(Received for publication, February 18, 1997, and in revised form, April 18, 1997)
From the Department of Biochemistry, We isolated a thiamin transporter gene,
THI10, from a genomic library of Saccharomyces
cerevisiae by the complementation of a yeast mutant defective in
thiamin transport activity. The THI10 gene contained an
open reading frame of 1,794 base pairs encoding a 598-amino acid
polypeptide with a calculated molecular weight of 66,903. The
nucleotide sequence of THI10 is completely identical to
that of an anonymous DNA (open reading frame L8083.2)
mapped to chromosome XII; two other genes (open reading frames
YOR071c and YOR192c) in chromosome XV are
extremely similar to THI10. Moreover, the THI10
gene product showed significant sequence homology with yeast allantoin
and uracil transporters. Hydropathy profile suggested that THI10
product is highly hydrophobic and contains many transmembrane regions.
Gene disruption of the THI10 locus completely abolished the
thiamin transport activity and thiamin binding activity in yeast plasma
membrane fraction. Both the transport and thiamin binding activities
were restored in the disrupted cells when the THI10 open
reading frame was expressed by yeast GAL1 promoter,
suggesting that the THI10 gene encodes for the thiamin
transport carrier protein. Northern blot analysis demonstrated that
THI10 gene expression is regulated at the mRNA level by
intracellular thiamin pyrophosphate and that it requires a positive
regulatory factor encoded by THI3 gene.
Volume 272, Number 31,
Issue of August 1, 1997
pp. 19165-19170
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
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