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Characterisation of the nascent polypeptide-associated complex in fission yeast

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Abstract

The nascent polypeptide-associated complex (NAC) is an abundant and phylogenetically conserved protein complex. It is composed of two subunits and interacts with nascent polypeptide chains emerging from the ribosome. It has been proposed to protect the nascent chains from premature interaction with other cell proteins, but has also been found to associate with DNA junctions, and to be involved in other processes including transcription regulation and mitochondrial protein import.

Here, we characterize NAC in fission yeast. We find that NAC is associated with ribosomes, while a significant fraction remains in a free form. The NAC alpha subunit contains a ubiquitin-associated (UBA) domain, which is found in several proteins involved in the ubiquitin–proteasome pathway for protein degradation. However, NAC does not associate with ubiquitin chains and mutants lacking NAC did not exhibit any obvious defects in protein degradation. Accordingly, we find that the NAC UBA domain belongs to an ancient and distinct subgroup of the UBA family. In contrast to the situation with budding yeast, fission yeast cells devoid of NAC were not temperature sensitive. However, they displayed resistance to the amino acid analogue canavanine, in accordance with the idea that NAC is involved in protein quality control.

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Abbreviations

GST:

Glutathione S-transferase

LDH:

Lactate dehydrogenase

NAC:

Nascent polypeptide-associated complex

SRP:

Signal recognition particle

UBA:

Ubiquitin-associated

References

  1. Wegrzyn RD, Deuerling E (2005) Molecular guardians for newborn proteins: ribosome-associated chaperones and their role in protein folding. Cell Mol Life Sci 62:2727–2738

    Article  PubMed  CAS  Google Scholar 

  2. Rospert S (2004) Ribosome function: governing the fate of a nascent polypeptide. Curr Biol 14:R386–R388

    Article  PubMed  CAS  Google Scholar 

  3. Rospert S, Dubaquie Y, Gautschi M (2002) Nascent-polypeptide-associated complex. Cell Mol Life Sci 59:1632–1639

    Article  PubMed  CAS  Google Scholar 

  4. Wiedmann B, Sakai H, Davis TA, Wiedmann M (1994) A protein complex required for signal-sequence-specific sorting and translocation. Nature 370:434–440

    Article  PubMed  CAS  Google Scholar 

  5. Bukau B, Deuerling E, Pfund C, Craig EA (2000) Getting newly synthesized proteins into shape. Cell 101:119–122

    Article  PubMed  CAS  Google Scholar 

  6. Grallath S, Schwarz JP, Bottcher UM, Bracher A, Hartl FU, Siegers K (2006) L25 functions as a conserved ribosomal docking site shared by nascent chain-associated complex and signal-recognition particle. EMBO Rep 7:78–84

    Article  PubMed  CAS  Google Scholar 

  7. Deng JM, Behringer RR (1995) An insertional mutation in the BTF3 transcription factor gene leads to an early postimplantation lethality in mice. Transgenic Res 4:264–269

    Article  PubMed  CAS  Google Scholar 

  8. Markesich DC, Gajewski KM, Nazimiec ME, Beckingham K (2000) Bicaudal encodes the Drosophila beta NAC homolog, a component of the ribosomal translational machinery. Development 127:559–572

    PubMed  CAS  Google Scholar 

  9. Wang S, Sakai H, Wiedmann M (1995) NAC covers ribosome-associated nascent chains thereby forming a protective environment for regions of nascent chains just emerging from the peptidyl transferase center. J Cell Biol 130:519–528

    Article  PubMed  CAS  Google Scholar 

  10. Möller I, Jung M, Beatrix B, Levy R, Kreibich G, Zimmermann R, Wiedmann M, Lauring B (1998) A general mechanism for regulation of access to the translocon: competition for a membrane attachment site on ribosomes. Proc Natl Acad Sci USA 95:13425–13430

    Article  PubMed  Google Scholar 

  11. Funfschilling U, Rospert S (1999) Nascent polypeptide-associated complex stimulates protein import into yeast mitochondria. Mol Biol Cell 10:3289–3299

    PubMed  CAS  Google Scholar 

  12. Whitby MC, Dixon J (2001) Fission yeast nascent polypeptide-associated complex binds to four-way DNA junctions. J Mol Biol 306:703–716

    Article  PubMed  CAS  Google Scholar 

  13. Quelo I, Akhouayri O, Prud’homme J, St-Arnaud R (2004) GSK3 beta-dependent phosphorylation of the alpha NAC coactivator regulates its nuclear translocation and proteasome-mediated degradation. Biochemistry 43:2906–2914

    Article  PubMed  CAS  Google Scholar 

  14. Franke J, Reimann B, Hartmann E, Köhler M, Wiedmann B (2001) Evidence for a nuclear passage of nascent polypeptide-associated complex subunits in yeast. J Cell Sci 117:2641–2648

    Google Scholar 

  15. Moreau A, Yotov WV, Glorieux FH, St-Arnaud R (1998) Bone-specific expression of the alpha chain of the nascent polypeptide-associated complex, a coactivator potentiating c-Jun-mediated transcription. Mol Cell Biol 18:1312–1321

    PubMed  CAS  Google Scholar 

  16. Yotov WV, Moreau A, St-Arnaud R (1998) The alpha chain of the nascent polypeptide-associated complex functions as a transcriptional coactivator. Mol Cell Biol 18:1303–1311

    PubMed  CAS  Google Scholar 

  17. Papachristou DJ, Batistatou A, Sykiotis GP, Varakis I, Papavassiliou AG (2003) Activation of the JNK-AP-1 signal transduction pathway is associated with pathogenesis and progression of human osteosarcomas. Bone 32:364–371

    Article  PubMed  CAS  Google Scholar 

  18. Yotov WV, St-Arnaud R (1996) Differential splicing-in of a proline-rich exon converts alphaNAC into a muscle-specific transcription factor. Genes Dev 10:1763–1772

    Article  PubMed  CAS  Google Scholar 

  19. Stilo R, Liguoro D, di Jeso B, Leonardi A, Vito P (2003) The alpha-chain of the nascent polypeptide-associated complex binds to and regulates FADD function. Biochem Biophys Res Commun 303:1034–1041

    Article  PubMed  CAS  Google Scholar 

  20. Zheng XM, Black D, Chambon P, Egly JM (1990) Sequencing and expression of complementary DNA for the general transcription factor BTF3. Nature 344:556–559

    Article  PubMed  CAS  Google Scholar 

  21. Andersen KM, Hofmann K, Hartmann-Petersen R (2005) Ubiquitin binding proteins: similar, but different. Essays Biochem 41:49–67

    PubMed  CAS  Google Scholar 

  22. Spreter T, Pech M, Beatrix B (2005) The crystal structure of archaeal nascent polypeptide-associated complex (NAC) reveals a unique fold and the presence of a ubiquitin-associated domain. J Biol Chem 280:15849–15854

    Article  PubMed  CAS  Google Scholar 

  23. Hofmann K, Bucher P (1996) The UBA domain: a sequence motif present in multiple enzyme classes of the ubiquitination pathway. Trends Biochem Sci 21:172–173

    Article  PubMed  CAS  Google Scholar 

  24. Glickman MH, Ciechanover A (2002) The ubiquitin-proteasome proteolytic pathway: destruction for the sake of construction. Physiol Rev 82:373–428

    PubMed  CAS  Google Scholar 

  25. Wilkinson CR, Seeger M, Hartmann-Petersen R, Stone M, Wallace M, Semple C, Gordon C (2001) Proteins containing the UBA domain are able to bind to multi-ubiquitin chains. Nat Cell Biol 3:939–943

    Article  PubMed  CAS  Google Scholar 

  26. Raasi S, Varadan R, Fushman D, Pickart C (2005) Diverse polyubiquitin interaction properties of ubiquitin-associated domains. Nat Struct Mol Biol 12:708–714

    Article  PubMed  CAS  Google Scholar 

  27. Moreno S, Klar A, Nurse P (1991) Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol 194:795–823

    Article  PubMed  CAS  Google Scholar 

  28. Bähler J, Wu JQ, Longtine MS, Shah NG, McKenzie A 3rd, Steever AB, Wach A, Philippsen P, Pringle JR (1998) Heterologous modules for efficient and versatile PCR-based gene targeting in Schizosaccharomyces pombe. Yeast 14:943–951

    Article  PubMed  Google Scholar 

  29. Cartwright P, Beilharz T, Hansen P, Garrett J, Lithgow T (1997) Mft52, an acid-bristle protein in the cytosol that delivers precursor proteins to yeast mitochondria. J Biol Chem 272:5320–5325

    Article  PubMed  CAS  Google Scholar 

  30. Shor B, Calaycay J, Rushbrook J, McLeod M (2003) Cpc2/RACK1 is a ribosome-associated protein that promotes efficient translation in Schizosaccharomyces pombe. J Biol Chem 278:49119–49128

    Article  PubMed  CAS  Google Scholar 

  31. Bergmeyer HU, Bernt E, Hess B (1963) Lactic dehydrogenase. In: Bergmeyer HU (ed) Methods of enzymatic analysis. Academic Press, Verlag Chemie GmbH, pp 736–743

  32. Heinemeyer W, Kleinschmidt JA, Saidowsky J, Escher C, Wolf DH (1991) Proteinase yscE, the yeast proteasome/multicatalytic-multifunctional proteinase: mutants unravel its function in stress induced proteolysis and uncover its necessity for cell survival. EMBO J 10:555–562

    PubMed  CAS  Google Scholar 

  33. Potashkin J, Wentz-Hunter K, Callaci J (1996) BTF3 is evolutionarily conserved in fission yeast. Biochim Biophys Acta 1308:182–184

    PubMed  Google Scholar 

  34. Reimann B, Bradsher J, Franke J, Hartmann E, Wiedmann M, Prehn S, Wiedmann B (1999) Initial characterization of the nascent polypeptide-associated complex in yeast. Yeast 15:397–407

    Article  PubMed  CAS  Google Scholar 

  35. Ellis RJ (2001) Macromolecular crowding: obvious but underappreciated. Trends Biochem Sci 26:597–604

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

The authors thank Ms A. Bonde Lauridsen for expert technical assistance, Dr M. C. Whitby for providing valuable reagents and Dr Klavs B. Hendil, Dr C. Gordon and Dr Peter S. Walmod for helpful discussions and comments on the manuscript. This work was funded by grants from the Lundbeck Foundation and the Danish Research Council to R.H.-P.

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Authors and Affiliations

  1. Institute for Molecular Biology and Physiology, University of Copenhagen, Universitetsparken 13, 2100, Copenhagen Ø, Denmark

    Katrine M. Andersen & Rasmus Hartmann-Petersen

  2. MRC Human Genetics Unit, Western General Hospital, Crewe Road, EH4 2XU, Edinburgh, Scotland, UK

    Colin A. Semple

Authors
  1. Katrine M. Andersen
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  2. Colin A. Semple
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  3. Rasmus Hartmann-Petersen
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Correspondence to Rasmus Hartmann-Petersen.

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Andersen, K.M., Semple, C.A. & Hartmann-Petersen, R. Characterisation of the nascent polypeptide-associated complex in fission yeast. Mol Biol Rep 34, 275–281 (2007). https://doi.org/10.1007/s11033-006-9043-5

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  • DOI: https://doi.org/10.1007/s11033-006-9043-5

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