Gly-46 and His-50 of yeast maltose transporter Mal21p are essential for its resistance against glucose-induced degradation.
The maltose transporter gene is situated at the MAL locus, which consists of genes for a transporter, maltase, and transcriptional activator. Five unlinked MAL loci (MAL1, MAL2, MAL3, MAL4, and MAL6) constitute a gene family in Saccharomyces cerevisiae. The expression of the maltose transporter is induced by maltose and repressed by glucose. The activity of the maltose transporter is also regulated post-translationally; Mal61p is rapidly internalized from the plasma membrane and degraded by ubiquitin-mediated proteolysis in the presence of glucose. We found that S. cerevisiae strain ATCC20598 harboring MAL21 could grow in maltose supplemented with a non- assimilable glucose analogue, 2-deoxyglucose, whereas strain ATCC96955 harboring MAL61 and strain CB11 with MAL31 and AGT1 could not. These observations implied a Mal21p-specific resistance against glucose-induced degradation. Mal21p found in ATCC20598 has 10 amino acids, including Gly-46 and His-50, that are inconsistent with the corresponding residues in Mal61p. The half-life of Mal21p for glucose-induced degradation was 118 min when expressed using the constitutive TPI1 promoter, which was significantly longer than that of Mal61p (25 min). Studies with mutant cells that are defective in endocytosis or the ubiquitination process indicated that Mal21p was less ubiquitinated than Mal61p, suggesting that Mal21p remains on the plasma membrane because of poor susceptibility to ubiquitination. Mutational studies revealed that both residues Gly-46 and His-50 in Mal21p are essential for the full resistance of maltose transporters against glucose-induced degradation.
Suntory Research Center, Osaka 618-8503, Japan. Haruyo_Hatanaka@suntory.co.jp
Haruyo
H
Hatanaka
Fumihiko
F
Omura
Yukiko
Y
Kodama
Toshihiko
T
Ashikari
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D017931:DNA Primers; D005947:Glucose / Q000494:pharmacology*; D005998:Glycine; D006639:Histidine; D008320:Maltose / Q000494:pharmacology; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D005810:Multigene Family; D016296:Mutagenesis; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000187:drug effects / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D027981:Symporters / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D015534:Trans-Activators / Q000378:metabolism
D017931:DNA Primers; D009004:Monosaccharide Transport Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D027981:Symporters; D015534:Trans-Activators; C067575:maltose transport system, S cerevisiae; D006639:Histidine; D008320:Maltose; D005947:Glucose; D005998:Glycine