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Fisheries Science

, Volume 71, Issue 2, pp 327–332 | Cite as

Expression of HSP70 in response to heat-shock and its cDNA cloning from Mediterranean blue mussel

  • Haruhiko Toyohara
  • Masatomi Hosoi
  • Isao Hayashi
  • Satoshi Kubota
  • Hisashi Hashimoto
  • Yoshihiro Yokoyama
Article

Abstract

Expression of HSP70 in response to heat-shock was investigated at the protein and mRNA levels in Mediterranean blue mussel. Western and Northern blot analyses revealed that HSP70 was expressed following heat-shock in the mantle at both protein and mRNA levels, suggesting that gene expression of HSP70 is implicated in the cellular response to heat-shock stress in mussel. It was then attempted to clone HSP70 cDNA in order to determine the primary structure of mussel HSP70. As a result, two full-length cDNA encoding HSP70 were isolated from a cDNA library prepared from the heat-shocked mantle. The isolated cDNA consist of single open reading frames of 2067 bp and 1911 bp which encode proteins of 689 amino acids and 637 amino acids, respectively. Both HSP70 cDNA encode an ATPase do main, and a substrate-binding do main in addition to a Glu-Glu-Val-Asp (EEVD) peptide motif that is specific for cytosolic HSP70. These findings suggest that the cDNA clones obtained in the present study encode cytosolic HSP70.

Key Words

heat-shock HSP70 mantle Mediterranean blue mussel Mytilus galloprovin-cialis stress 

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References

  1. 1.
    Kültz D. Evolution of the cellular stress proteome: from monophyletic origin to ubiquitous function. J. Exp. Biol. 2003; 206: 3119–3124PubMedCrossRefGoogle Scholar
  2. 2.
    Morimoto RI, Tissieres A, Georgopoulos C. Stress Protein in Biology and Medicine, Cold Spring Harbour Laboratory Press, New York, 1994.Google Scholar
  3. 3.
    Somero GN. Proteins and temperature. Annu. Rev. Physiol. 1995: 57: 43–68.PubMedCrossRefGoogle Scholar
  4. 4.
    Somero GN, Hofman GE. Temperature thresholds for protein adaptation: when does temperaturt change start to ‘hurt’? In: Wood CM, McDonald DG (eds). Global Warming: Implications for Freshwater and Marine Fish. Cambridge University Press, Cambridge, 1996; 1–24Google Scholar
  5. 5.
    Margulis BA, Antropova OY, Kharazova AD. 70 kDa heat shock proteins from mollusk and human cells have common structural and functional domains. Comp. Biochem. Physiol. B 1989; 94: 621–623.PubMedCrossRefGoogle Scholar
  6. 6.
    Veldhuizen-Tsoerkan MB, Howerda DA, Zandee M. Synthesis of stress proteins under normal and heat shock conditions in gill tissue of sea mussed (Mytilus edulis) after chronic exposure to cadmium. Comp. Biochem. Physiol. C 1991: 100: 699–706.PubMedCrossRefGoogle Scholar
  7. 7.
    Veldhuizen-Tsoerkan MB, Holwerda DA, Bont AM, Smaal AC, Zandee DI. A field study on stress indices in the sea mussel, Mytilus edulis: application of the ‘stress approach’ in biomonitoring. Arch. Environ. Contam. Toxicol. 1991; 21: 497–504.PubMedCrossRefGoogle Scholar
  8. 8.
    Sanders BM, Pascoe VM, Nakagawa PA, Martin LS. Persistence of the heat-shock response over time in a common Mytilus mussel. Mol. Mar. Biol. Biotech. 1992; 1: 147–154.Google Scholar
  9. 9.
    Hofmann GE, Somero GN. Evidence for protein damage at environmental temperature. Seasonal changes levels of ubiquitin conjugates and HSP70 in the intertidal mussel Mytilus trossulus. J. Exp. Zool. 1995; 198: 1509–1518.Google Scholar
  10. 10.
    Smerdon GR, Chapple JP, Hawkins AJS. The simultaneous immunological detection of four stress-70 protein isoforms in Mytilus edulis. Mar. Rnviron. Res. 1995; 40: 399–407.CrossRefGoogle Scholar
  11. 11.
    Hofmann GE, Somero GN. Protein ubiquitination and stress protein synthesis in Mytilus trossulus occurs during recovery from tidal emersion. Mol. Mar. Biol. Biotech. 1996; 5: 175–184.Google Scholar
  12. 12.
    Minier C, Borghi VV, Moore MN, Porte C. Seasonal variation of MXR and stress proteins in the common mussel, Mytilus galloprovincialis. Aquat. Toxicol. 2000; 50: 167–176.PubMedCrossRefGoogle Scholar
  13. 13.
    Porte C, Biosca X, Sole M, Albaiges J. The integrated use of chemical analysis, cytochrome P450 and stress proteins in mussels to assess pollution along the Galician coast (NW Spain). Environ. Pollut. 2001; 112: 261–268.PubMedCrossRefGoogle Scholar
  14. 14.
    Snyder MJ, Girvetz E, Mulder EP. Induction of marine molluscs stress proteins by chemical or physical stress. Arch. Environ. Contam. Toxicol. 2001; 41: 22–29.PubMedCrossRefGoogle Scholar
  15. 15.
    Buckley BA, Owen ME, Hofmann GE. Adjusting the thermostat: the threshold induction temperature for the heatshock response in the intertidal mussels (genus Mytilus) changes as a function of thermal history. J. Exp. Biol. 2001; 204: 3571–3579.PubMedGoogle Scholar
  16. 16.
    Helmuth BS, Hofmann GE. Microhabitats, thermal heterogeneity, and patterns of physiological stress in the rocky intertidal zone. Biol. Bull. 2001; 201: 374–384.PubMedCrossRefGoogle Scholar
  17. 17.
    Randlowska M, Pempkowiak J. Stress-70 as indicator of heavy metal accumulation in the blue mussel Mytilus edulis. Environ. Int. 2002; 27: 605–608.CrossRefGoogle Scholar
  18. 18.
    Lyons C, Dowling V, Tedengren M, Gardestrom J, Hartl MG, O’Brien N, van Pelt FN, O’Halloran J, Sheehan D. Variability of heat shock proteins and glutathione S-transferase in gill and digestive gland of blue mussel, Mytilus edulis. Mar. Environ. Res. 2003; 56: 585–597.PubMedCrossRefGoogle Scholar
  19. 19.
    Malagoli D, Lusvardi M, Gobba F, Ottaviani E. 50 HZ magnetic fields activate mussel immunocyte p38 MAP kinase and induce HSP70 and 90. Comp. Biochem. Physiol. C 2004; 137: 75–79.CrossRefGoogle Scholar
  20. 20.
    Sambrook J, Russel DW. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, New York, 2001.Google Scholar
  21. 21.
    Yamashita M, Hiryoshi K, Nagata K. Characterization of multiple members of the HSP70 family in platyfish culture cells: molecular evolution of stress protein HSP70 in vertebrates. Gene 2004; 336: 207–218.PubMedCrossRefGoogle Scholar
  22. 22.
    Kozak M. Initiation of translation in prokaryotes and eukaryotes. Gene 1999; 234: 187–208.PubMedCrossRefGoogle Scholar

Copyright information

© The Japanese Society of Fisheries Science 2005

Authors and Affiliations

  • Haruhiko Toyohara
    • 1
  • Masatomi Hosoi
    • 1
  • Isao Hayashi
    • 1
  • Satoshi Kubota
    • 2
  • Hisashi Hashimoto
    • 3
  • Yoshihiro Yokoyama
    • 4
  1. 1.Division of Applied Biosciences, Graduate School of AgricultureKyoto UniversityKyotoJapan
  2. 2.Division of Human Health and Medical Science, Graduate School of Kuroshio ScienceKochi UniversityNankokuJapan
  3. 3.Bioscience and Biotechnology CenterNagoya UniversityNagoyaJapan
  4. 4.Department of Marine BioscienceFukui Prefectural UniversityFukuiJapan

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