Heterologous expression of metabotropic glutamate receptor subtype 1 in Saccharomyces cerevisiae.
The upstream region of the isocitrate lyase gene (UPR-ICL) from the n-alkane-utilizing yeast Candida tropicalis serves as a useful promoter of gene expression in the yeast Saccharomyces cerevisiae. The production of rat metabotropic glutamate receptor 1alpha (mGluR1alpha), which belongs to the G-protein-coupled receptor (GPCR) family, was tested under the control of UPR-ICL. Expression of mGluR1alpha was found in recombinant clones and enhanced by replacing the signal sequence of mGluR1alpha with the corresponding region of the alpha-factor receptor (Ste2), which is a GPCR found in S. cerevisiae. Moreover, the membrane fraction from a recombinant clone associated with Vesl-1S/Homer-1a protein binds the mGluR1alpha in rat cerebellum. These results suggest that the UPR-ICL-controlled gene expression system is useful for heterologous GPCRs in S. cerevisiae.
Sugiyama
K
K
GenCom, 11 Minamiooya, Machida 194-8511, Japan. sugiyama@nihs.go.jp
Niki
T P
TP
Inokuchi
K
K
Teranishi
Y
Y
Ueda
M
M
Tanaka
A
A
D000818:Animals; D041022:Candida tropicalis / Q000235:genetics; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D003001:Cloning, Molecular; D007522:Isocitrate Lyase / Q000235:genetics; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals / Q000235:genetics; D051381:Rats; D018094:Receptors, Metabotropic Glutamate / Q000096:biosynthesis* / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism
D021382:Protein Sorting Signals; D018094:Receptors, Metabotropic Glutamate; D011993:Recombinant Fusion Proteins; C104077:metabotropic glutamate receptor type 1; D007522:Isocitrate Lyase