Skip to main content
SpringerLink
Log in

Characterization of Ire1 in the yeast Yarrowia lipolytica reveals an important role for the Sls1 nucleotide exchange factor in unfolded protein response regulation

  • Research Article
  • Published:
Current Genetics Aims and scope Submit manuscript

Abstract

Following endoplasmic reticulum (ER) stress, eukaryotic cells trigger a conserved signal transduction pathway called the unfolded protein response (UPR) that regulates the ER’s capacity to perform protein folding according to cellular demand. In Saccharomyces cerevisiae, the UPR is initiated by Ire1, a type I transmembrane serine/threonine kinase/endoribonuclease, that senses unfolded protein levels within the ER in collaboration with the ER Hsp70-family member, BiP/Kar2. Here, we report on the characterization of the Yarrowia lipolytica Ire1 ortholog. Our results show that Sls1, a nucleotide exchange factor for BiP, has important functions in regulating ER stress and the interaction of BiP and Ire1. They suggest that Sls1 regulates this interaction, by stimulating the conversion of BiP from the ADP-bound to the ATP-bound state, which favors its interaction with Ire1. Moreover, we identified known and new partners for Ire1 using the Tandem Affinity Purification (TAP) approach.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Anttonen AK, Mahjneh I, Hamalainen RH, Lagier-Tourenne C, Kopra O, Waris L, Anttonen M, Joensuu T, Kalimo H, Paetau A, Tranebjaerg L, Chaigne D, Koenig M, Eeg-Olofsson O, Udd B, Somer M, Somer H, Lehesjoki AE (2005) The gene disrupted in Marinesco–Sjogren syndrome encodes SIL1, an HSPA5 cochaperone. Nat Genet 37:1309–1311

    Article  PubMed  CAS  Google Scholar 

  • Babour A, Beckerich JM, Gaillardin C (2004) Identification of an UDP-Glc:glycoprotein glucosyltransferase in the yeast Yarrowia lipolytica. Yeast 21:11–24

    Article  PubMed  CAS  Google Scholar 

  • Bertolotti A, Zhang Y, Hendershot LM, Harding HP, Ron D (2000) Dynamic interaction of BiP and ER stress transducers in the unfolded-protein response. Nat Cell Biol 2:326–332

    Article  PubMed  CAS  Google Scholar 

  • Bicknell AA, Babour A, Federovitch CM, Niwa M (2007) A novel role in cytokinesis reveals a housekeeping function for the unfolded protein response. J Cell Biol 177:1017–1027

    Article  PubMed  CAS  Google Scholar 

  • Boisrame A, Beckerich JM, Gaillardin C (1996) Sls1p, an endoplasmic reticulum component, is involved in the protein translocation process in the yeast Yarrowia lipolytica. J Biol Chem 271:11668–11675

    Article  PubMed  CAS  Google Scholar 

  • Boisrame A, Kabani M, Beckerich JM, Hartmann E, Gaillardin C (1998) Interaction of Kar2p and Sls1p is required for efficient co-translational translocation of secreted proteins in the yeast Yarrowia lipolytica. J Biol Chem 273:30903–30908

    Article  PubMed  CAS  Google Scholar 

  • Borradaile NM, Buhman KK, Listenberger LL, Magee CJ, Morimoto ET, Ory DS, Schaffer JE (2006) A critical role for eukaryotic elongation factor 1A–1 in lipotoxic cell death. Mol Biol Cell 17:770–778

    Article  PubMed  CAS  Google Scholar 

  • Chuang SM, Chen L, Lambertson D, Anand M, Kinzy TG, Madura K (2005) Proteasome-mediated degradation of cotranslationally damaged proteins involves translation elongation factor 1A. Mol Cell Biol 25:403–413

    Article  PubMed  CAS  Google Scholar 

  • Chung KT, Shen Y, Hendershot LM (2002) BAP, a mammalian BiP-associated protein, is a nucleotide exchange factor that regulates the ATPase activity of BiP. J Biol Chem 277:47557–47563

    Article  PubMed  CAS  Google Scholar 

  • Cox JS, Shamu CE, Walter P (1993) Transcriptional induction of genes encoding endoplasmic reticulum resident proteins requires a transmembrane protein kinase. Cell 73:1197–1206

    Article  PubMed  CAS  Google Scholar 

  • Cox JS, Walter P (1996) A novel mechanism for regulating activity of a transcription factor that controls the unfolded protein response. Cell 87:391–404

    Article  PubMed  CAS  Google Scholar 

  • Credle JJ, Finer-Moore JS, Papa FR, Stroud RM, Walter P (2005) On the mechanism of sensing unfolded protein in the endoplasmic reticulum. Proc Natl Acad Sci USA 102:18773–18784

    Article  PubMed  CAS  Google Scholar 

  • Gonen H, Smith CE, Siegel NR, Kahana C, Merrick WC, Chakraburtty K, Schwartz AL, Ciechanover A (1994) Protein synthesis elongation factor EF-1 alpha is essential for ubiquitin-dependent degradation of certain N alpha-acetylated proteins and may be substituted for by the bacterial elongation factor EF-Tu. Proc Natl Acad Sci USA 91:7648–7652

    Article  PubMed  CAS  Google Scholar 

  • Grant PA, Schieltz D, Pray-Grant MG, Yates JR 3rd, Workman JL (1998) The ATM-related cofactor Tra1 is a component of the purified SAGA complex. Mol Cell 2:863–867

    Article  PubMed  CAS  Google Scholar 

  • Harding HP, Zhang Y, Ron D (1999) Protein translation and folding are coupled by an endoplasmic-reticulum-resident kinase. Nature 397:271–274

    Article  PubMed  CAS  Google Scholar 

  • Haze K, Yoshida H, Yanagi H, Yura T, Mori K (1999) Mammalian transcription factor ATF6 is synthesized as a transmembrane protein and activated by proteolysis in response to endoplasmic reticulum stress. Mol Biol Cell 10:3787–3799

    PubMed  CAS  Google Scholar 

  • Hotokezaka Y, Tobben U, Hotokezaka H, Van Leyen K, Beatrix B, Smith DH, Nakamura T, Wiedmann M (2002) Interaction of the eukaryotic elongation factor 1A with newly synthesized polypeptides. J Biol Chem 277:18545–18551

    Article  PubMed  CAS  Google Scholar 

  • Kabani M, Beckerich JM, Brodsky JL (2002a) Nucleotide exchange factor for the yeast Hsp70 molecular chaperone Ssa1p. Mol Cell Biol 22:4677–4689

    Article  PubMed  CAS  Google Scholar 

  • Kabani M, Beckerich JM, Brodsky JL (2003) The yeast Sls1 and Fes1p proteins define a new family of Hsp70 nucleotide exchange factors. Curr Gen 4:465–473

    Article  CAS  Google Scholar 

  • Kabani M, Beckerich JM, Gaillardin C (2000a) Sls1p stimulates Sec63p-mediated activation of Kar2p in a conformation-dependent manner in the yeast endoplasmic reticulum. Mol Cell Biol 20:6923–6934

    Article  PubMed  CAS  Google Scholar 

  • Kabani M, Boisrame A, Beckerich JM, Gaillardin C (2000b) A highly representative two-hybrid genomic library for the yeast Yarrowia lipolytica. Gene 241:309–315

    Article  PubMed  CAS  Google Scholar 

  • Kabani M, McLellan C, Raynes DA, Guerriero V, Brodsky JL (2002b) HspBP1, a homologue of the yeast Fes1 and Sls1 proteins, is an Hsc70 nucleotide exchange factor. FEBS Lett 531:339–342

    Article  PubMed  CAS  Google Scholar 

  • Kimata Y, Kimata YI, Shimizu Y, Abe H, Farcasanu IC, Takeuchi M, Rose MD, Kohno K (2003) Genetic evidence for a role of BiP/Kar2 that regulates Ire1 in response to accumulation of unfolded proteins. Mol Biol Cell 14:2559–2569

    Article  PubMed  CAS  Google Scholar 

  • Kimata Y, Oikawa D, Shimizu Y, Ishiwata-Kimata Y, Kohno K (2004) A role for BiP as an adjustor for the endoplasmic reticulum stress-sensing protein Ire1. J Cell Biol 167:445–456

    Article  PubMed  CAS  Google Scholar 

  • Kippert F (1995) A rapid permeabilization procedure for accurate quantitative determination of beta-galactosidase activity in yeast cells. FEMS Microbiol Lett 128:201–206

    PubMed  CAS  Google Scholar 

  • Liu CY, Wong HN, Schauerte JA, Kaufman RJ (2002) The protein kinase/endoribonuclease IRE1alpha that signals the unfolded protein response has a luminal N-terminal ligand-independent dimerization domain. J Biol Chem 277:18346–18356

    Article  PubMed  CAS  Google Scholar 

  • Liu CY, Xu Z, Kaufman RJ (2003) Structure and intermolecular interactions of the luminal dimerization domain of human IRE1alpha. J Biol Chem 278:17680–17687

    Article  PubMed  CAS  Google Scholar 

  • Marcu MG, Doyle M, Bertolotti A, Ron D, Hendershot L, Neckers L (2002) Heat shock protein 90 modulates the unfolded protein response by stabilizing IRE1alpha. Mol Cell Biol 22:8506–8513

    Article  PubMed  CAS  Google Scholar 

  • Mori K, Ma W, Gething MJ, Sambrook J (1993) A transmembrane protein with a cdc2+/CDC28-related kinase activity is required for signaling from the ER to the nucleus. Cell 74:743–756

    Article  PubMed  CAS  Google Scholar 

  • Okamura K, Kimata Y, Higashio H, Tsuru A, Kohno K (2000) Dissociation of Kar2p/BiP from an ER sensory molecule, Ire1p, triggers the unfolded protein response in yeast. Biochem Biophys Res Commun 279:445–450

    Article  PubMed  CAS  Google Scholar 

  • Patil C, Walter P (2001) Intracellular signaling from the endoplasmic reticulum to the nucleus: the unfolded protein response in yeast and mammals. Curr Opin Cell Biol 13:349–355

    Article  PubMed  CAS  Google Scholar 

  • Rigaut G, Shevchenko A, Rutz B, Wilm M, Mann M, Seraphin B (1999) A generic protein purification method for protein complex characterization and proteome exploration. Nat Biotechnol 17:1030–1032

    Article  PubMed  CAS  Google Scholar 

  • Rothstein R (1991) Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast. Methods Enzymol 194:281–301

    Article  PubMed  CAS  Google Scholar 

  • Schroder M, Kaufman RJ (2005) The mammalian unfolded protein response. Annu Rev Biochem 74:739–789

    Article  PubMed  CAS  Google Scholar 

  • Senderek J, Krieger M, Stendel C, Bergmann C, Moser M, Breitbach-Faller N, Rudnik-Schoneborn S, Blaschek A, Wolf NI, Harting I, North K, Smith J, Muntoni F, Brockington M, Quijano-Roy S, Renault F, Herrmann R, Hendershot LM, Schroder JM, Lochmuller H, Topaloglu H, Voit T, Weis J, Ebinger F, Zerres K (2005) Mutations in SIL1 cause Marinesco-Sjogren syndrome, a cerebellar ataxia with cataract and myopathy. Nat Genet 37:1312–1314

    Article  PubMed  CAS  Google Scholar 

  • Shamu CE, Walter P (1996) Oligomerization and phosphorylation of the Ire1p kinase during intracellular signaling from the endoplasmic reticulum to the nucleus. EMBO J 15:3028–3039

    PubMed  CAS  Google Scholar 

  • Tirasophon W, Welihinda AA, Kaufman RJ (1998) A stress response pathway from the endoplasmic reticulum to the nucleus requires a novel bifunctional protein kinase/endoribonuclease (Ire1p) in mammalian cells. Genes Dev 12:1812–1824

    Article  PubMed  CAS  Google Scholar 

  • Todd-Corlett A, Jones E, Seghers C, Gething MJ (2007) Lobe IB of the ATPase domain of Kar2p/BiP interacts with Ire1p to negatively regulate the unfolded protein response in Saccharomyces cerevisiae. J Mol Biol 367:770–787

    Article  PubMed  CAS  Google Scholar 

  • Travers KJ, Patil CK, Wodicka L, Lockhart DJ, Weissman JS, Walter P (2000) Functional and genomic analyses reveal an essential coordination between the unfolded protein response and ER-associated degradation. Cell 101:249–258

    Article  PubMed  CAS  Google Scholar 

  • Tyson JR, Stirling CJ (2000) LHS1 and SIL1 provide a lumenal function that is essential for protein translocation into the endoplasmic reticulum. Embo J 19:6440–6452

    Article  PubMed  CAS  Google Scholar 

  • Wang XZ, Harding HP, Zhang Y, Jolicoeur EM, Kuroda M, Ron D (1998) Cloning of mammalian Ire1 reveals diversity in the ER stress responses. Embo J 17:5708–5717

    Article  PubMed  CAS  Google Scholar 

  • Welihinda AA, Tirasophon W, Green SR, Kaufman RJ (1997) Gene induction in response to unfolded protein in the endoplasmic reticulum is mediated through Ire1p kinase interaction with a transcriptional coactivator complex containing Ada5p. Proc Natl Acad Sci USA 94:4289–4294

    Article  PubMed  CAS  Google Scholar 

  • Welihinda AA, Tirasophon W, Kaufman RJ (2000) The transcriptional co-activator ADA5 is required for HAC1 mRNA processing in vivo. J Biol Chem 275:3377–3381

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Author notes

  1. Anna Babour

    Present address: University of California, San Diego, NSB 5315 9500 Gilman Drive, La Jolla, CA, 92093, USA

  2. Mehdi Kabani

    Present address: Laboratoire d’Enzymologie et Biochimie Structurales (LEBS), CNRS, 91198, Gif-sur-Yvette, France

Authors and Affiliations

  1. Laboratoire de Microbiologie et Génétique moléculaire, INRA, CNRS, Institut National Agronomique Paris-Grignon, 78850, Thiverval-Grignon, France

    Anna Babour, Mehdi Kabani, Anita Boisramé & Jean-Marie Beckerich

Authors
  1. Anna Babour
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mehdi Kabani
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anita Boisramé
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jean-Marie Beckerich
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Anna Babour.

Additional information

Communicated by K. Kuchler.

Electronic supplementary material

Below is the link to the electronic supplementary material.

294_2008_190_MOESM1_ESM.doc

Sequence of YlIRE1. The complete sequence of YlIRE1 is given together with the ORF translation. The back arrow shows the putative signal sequence cleavage site. The black rectangle depicts the transmembrane segment. (DOC 48 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Babour, A., Kabani, M., Boisramé, A. et al. Characterization of Ire1 in the yeast Yarrowia lipolytica reveals an important role for the Sls1 nucleotide exchange factor in unfolded protein response regulation. Curr Genet 53, 337–346 (2008). https://doi.org/10.1007/s00294-008-0190-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00294-008-0190-1

Keywords

Search

Navigation