The calnexin homologue cnx1+ in Schizosaccharomyces pombe, is an essential gene which can be complemented by its soluble ER domain.
Secretory proteins become folded by the action of a number of molecular chaperones soon after they enter the endoplasmic reticulum (ER). In mammalian cells, the ER membrane protein calnexin has been shown to be a molecular chaperone involved in the folding of secretory proteins and in the assembly of cell surface receptor complexes. We have used a PCR strategy to identify the Schizosaccharomyces pombe calnexin homologue, cnx1+. The cnx1+ encoded protein, Cnx1, was shown to be a calcium binding type I integral membrane glycoprotein. At its 5' end, the cnx1+ gene has consensus heat shock transcriptional control elements and was inducible by heat shock and by the calcium ionophore A23187. Unlike the sequence-related Saccharomyces cerevisiae CNE1 gene, the S.pombe cnx1+ gene was essential for cell viability. The full-length Cnx1 protein was able to complement the cnx1+ gene disruption but the full-length mammalian calnexin could not. The ER lumenal domain of Cnx1, which was secreted from cells, was capable of complementing the cnx1::ura4 lethal phenotype. The equivalent region of mammalian calnexin has been shown to possess molecular chaperone activity. It is possible that the lethal phenotype is caused by the absence of this chaperone activity in the S.pombe cnx1+ gene disruption.
Parlati
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F
Department of Biology, McGill University, Montreal, Quebec, Canada.
Dignard
D
D
Bergeron
J J
JJ
Thomas
D Y
DY
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*; D037281:Calnexin; D004285:Dogs; D004721:Endoplasmic Reticulum / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D018832:Molecular Chaperones / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D012568:Schizosaccharomyces / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D014498:Uracil / Q000096:biosynthesis
D002135:Calcium-Binding Proteins; C094757:Cnx1 protein, S pombe; D006360:Heat-Shock Proteins; D008562:Membrane Glycoproteins; D018832:Molecular Chaperones; D037281:Calnexin; D014498:Uracil