Expression of the four subunits of the Torpedo californica nicotinic acetylcholine receptor in Saccharomyces cerevisiae.
Yeast expression vectors were constructed containing complementary DNA encoding the alpha-, beta-, gamma-, and delta-subunits of the Torpedo californica nicotinic acetylcholine receptor under the control of the Saccharomyces cerevisiae alcohol dehydrogenase promoter. All four plasmids were integrated into the yeast genome of a single yeast cell. The resulting yeast strain synthesized polypeptides novel to yeast that had the molecular weights and antigenic properties similar to the authentic T. californica receptor alpha-, gamma, and delta-subunits. The beta-subunit polypeptide could not be detected in this yeast strain, even though the poly(A)+ RNA from this strain contained all the information necessary for the expression of functional acetylcholine receptors in Xenopus laevis oocytes. The replacement of the beta-subunit mRNA 5'-untranslated leader and its N-terminal signal sequence by the corresponding alpha-subunit sequences, however, resulted in the expression of the beta-subunit polypeptide in yeast grown at 5 degrees C.
Jansen
K U
KU
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853.
Conroy
W G
WG
Claudio
T
T
Fox
T D
TD
Fujita
N
N
Hamill
O
O
Lindstrom
J M
JM
Luther
M
M
Nelson
N
N
Ryan
K A
KA
D000818:Animals; D001483:Base Sequence; D004271:DNA, Fungal / Q000235:genetics; D005800:Genes, Fungal; D005813:Genes, Synthetic; D005822:Genetic Vectors; D008564:Membrane Potentials; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D011061:Poly A / Q000502:physiology; D012331:RNA, Fungal / Q000502:physiology; D011978:Receptors, Nicotinic / Q000096:biosynthesis / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D014101:Torpedo / Q000235:genetics / Q000378:metabolism*; D014982:Xenopus laevis
D004271:DNA, Fungal; D012331:RNA, Fungal; D011978:Receptors, Nicotinic; D011061:Poly A