title: A role for the DnaJ homologue Scj1p in protein folding in the yeast endoplasmic reticulum.
abstract: Members of the eukaryotic heat shock protein 70 family (Hsp70s) are regulated by protein cofactors that contain domains homologous to bacterial DnaJ. Of the three DnaJ homologues in the yeast rough endoplasmic reticulum (RER; Scj1p, Sec63p, and Jem1p), Scj1p is most closely related to DnaJ, hence it is a probable cofactor for Kar2p, the major Hsp70 in the yeast RER. However, the physiological role of Scj1p has remained obscure due to the lack of an obvious defect in Kar2p-mediated pathways in scj1 null mutants. Here, we show that the Deltascj1 mutant is hypersensitive to tunicamycin or mutations that reduce N-linked glycosylation of proteins. Although maturation of glycosylated carboxypeptidase Y occurs with wild-type kinetics in Deltascj1 cells, the transport rate for an unglycosylated mutant carboxypeptidase Y (CPY) is markedly reduced. Loss of Scj1p induces the unfolded protein response pathway, and results in a cell wall defect when combined with an oligosaccharyltransferase mutation. The combined loss of both Scj1p and Jem1p exaggerates the sensitivity to hypoglycosylation stress, leads to further induction of the unfolded protein response pathway, and drastically delays maturation of an unglycosylated reporter protein in the RER. We propose that the major role for Scj1p is to cooperate with Kar2p to mediate maturation of proteins in the RER lumen.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01655-0103, USA.
authors.forename: S
authors.initials: S
authors.lastname: Silberstein
authors.forename: G
authors.initials: G
authors.lastname: Schlenstedt
authors.forename: P A
authors.initials: PA
authors.lastname: Silver
authors.forename: R
authors.initials: R
authors.lastname: Gilmore
mesh_terms: D000483:Alleles; D018833:Chaperonins / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000737:chemistry / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D006031:Glycosylation; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018832:Molecular Chaperones; D016296:Mutagenesis / Q000502:physiology; D009844:Oligosaccharides / Q000378:metabolism; D018384:Oxidative Stress / Q000502:physiology; D017510:Protein Folding; D012333:RNA, Messenger / Q000032:analysis; D029701:Saccharomyces cerevisiae Proteins; D013696:Temperature; D015003:Yeasts / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D005656:Fungal Proteins; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; C106168:JEM1 protein, S cerevisiae; C060153:KAR2 protein, yeast; D008565:Membrane Proteins; D018832:Molecular Chaperones; D009844:Oligosaccharides; D012333:RNA, Messenger; C067634:SCJ1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D018833:Chaperonins