title: Activation of the Ras-cAMP signal transduction pathway inhibits the proteasome-independent degradation of misfolded protein aggregates in the endoplasmic reticulum lumen.
abstract: Many kinds of misfolded secretory proteins are known to be degraded in the endoplasmic reticulum (ER). Dislocation of misfolded proteins from the ER to the cytosol and subsequent degradation by the proteasome have been demonstrated. Using the yeast Saccharomyces cerevisiae, we have been studying the secretion of a heterologous protein, Rhizopus niveus aspartic proteinase-I (RNAP-I). Previously, we found that the pro sequence of RNAP-I is important for the folding and secretion, and that Deltapro, a mutated derivative of RNAP-I in which the entire region of the pro sequence is deleted, forms gross aggregates in the yeast ER. In this study, we show that the degradation of Deltapro occurs independently of the proteasome. Its degradation was not inhibited either by a potent proteasome inhibitor or in a proteasome mutant. We also show that neither the export from the ER nor the vacuolar proteinase is required for the degradation of Deltapro. These results raise the possibility that the Deltapro aggregates are degraded in the ER lumen. We have isolated a yeast mutant in which the degradation of Deltapro is delayed. We show that the mutated gene is IRA2, which encodes a GTPase-activating protein for Ras. Because Ira2 protein is a negative regulator of the Ras-cAMP pathway, this result suggests that hyperactivation of the Ras-cAMP pathway inhibits the degradation of Deltapro. Consistently, down-regulation of the Ras-cAMP pathway in the ira2 mutant suppressed the defect of the degradation of Deltapro. Thus, the Ras-cAMP signal transduction pathway seems to control the proteasome-independent degradation of the ER misfolded protein aggregates.
authors.affiliation: Department of Biotechnology, The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. kyohei@postman.riken.go.jp
authors.forename: K
authors.initials: K
authors.lastname: Umebayashi
authors.forename: R
authors.initials: R
authors.lastname: Fukuda
authors.forename: A
authors.initials: A
authors.lastname: Hirata
authors.forename: H
authors.initials: H
authors.lastname: Horiuchi
authors.forename: A
authors.initials: A
authors.lastname: Nakano
authors.forename: A
authors.initials: A
authors.lastname: Ohta
authors.forename: M
authors.initials: M
authors.lastname: Takagi
mesh_terms: D016282:Aspartic Acid Endopeptidases / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000648:ultrastructure; D006868:Hydrolysis; D008854:Microscopy, Electron; D009097:Multienzyme Complexes / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000235:genetics; D015398:Signal Transduction; D018631:ras Proteins / Q000378:metabolism*
chemical_list: D009097:Multienzyme Complexes; D000242:Cyclic AMP; D003546:Cysteine Endopeptidases; D016282:Aspartic Acid Endopeptidases; C100354:aspartic proteinase-I; D046988:Proteasome Endopeptidase Complex; D018631:ras Proteins