Chinese Science Bulletin

, Volume 49, Issue 22, pp 2385–2393 | Cite as

Cloning, sequencing and expression analysis of cDNA encoding a constitutive heat shock protein 70 (HSC70) inFenneropenaeus chinensis

  • Chuanzhen Jiao
  • Zaizhao Wang
  • Fuhua Li
  • Chengsong Zhang
  • Jianhai Xiang
Articles
Received:
Accepted:
  • 27 Downloads

Abstract

The cDNA encodinghsc70 of Chinese shrimpFenneropenaeus chinensis was cloned from hepatopancreas by RT-PCR based on its EST sequence. The full length cDNA of 2090 bp contained an open reading frame of 1956 nucleotides and partial 5′- and 3′-untranslated region(5′- and 3′-UTR). PCR amplification and sequencing analysis showed the existence of introns in the region of 1–547 bp, but they did not exist in the region of 548–2090 bp ofhsc70 cDNA. When the deduced 652 amino acid sequence of HSC70 was compared with the members of HSP70 family from other organisms, the results showed 85.9% similarity with HSC71 fromOncorhynchus mykiss and HSC70 fromHomo sapiens. It also exhibited 85.8% similarity with HSP70 fromMus musculu and 85.4% with HSC70 fromManduca sexta. Expression analysis showed that hsc70 mRNA was espressed constitutively in hepatopancreas, muscle, eyestalks, haemocytes, heart, ovary, intestine and gills inFenneropenaeus chinensis. No difference could be detected onhsc70 mRNA level in muscle between heat-shocked and control animals.

Keywords

Fenneropenaeus chinensis HSC70 cloning expression 
This is a preview of subscription content, log in to check access.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Hightower, L. E., Heat shock, stress proteins, chaperones and proteotoxicity, Cell, 1991, 66: 191–197.CrossRefGoogle Scholar
  2. 2.
    Hendrick, J. P., Hartl, F. U., Molecular chaperone functions of heat shock proteins, Annu. Rev. Biochem., 1993, 62: 349–384.CrossRefGoogle Scholar
  3. 3.
    Welch, W. J., How cells respond to stress, Sci. Am., 1993, 269: 56–64.CrossRefGoogle Scholar
  4. 4.
    Basu, N., Todghama, A. E., Ackerman, P. A., Heat shock protein genes and their functional significance in fish, Gene, 2002, 295: 173–183.CrossRefGoogle Scholar
  5. 5.
    Juliann, G. K., George, C. T., Heat Shock Protein 70 kD: Molecular Biology, Biochemistry and Physiology, Pharmacol. Ther., 1998, 80(2): 183–201.CrossRefGoogle Scholar
  6. 6.
    Parsell, D. A., Lindquist, S., The function of heat-shock proteins in stress tolerance: Degradation and reactivation of damaged proteins, Annu. Rev. Genet., 1993, 27: 437–496.CrossRefGoogle Scholar
  7. 7.
    Morimoto, R. I., Santoro, M. G., Stress-inducible responses and heat shock proteins: New pharmacological targets for cytoprotection, Nat. Biotechnol., 1998, 16: 833–838.CrossRefGoogle Scholar
  8. 8.
    Jindal, S., Markovsky, M., Stress responses to viral infection, Trends Microbiol., 1994, 2 (3): 89–91.CrossRefGoogle Scholar
  9. 9.
    Sagara, J., Kawai, A., Identification of heat shock protein70 in the rabies virion, Virology, 1992, 190 (2): 845–848.CrossRefGoogle Scholar
  10. 10.
    Carlos, A. G., Daniela, B., Selene, Z., Heat shock cognate protein 70 is involved in rotavirus cell entry, J. Virol., 2002, 76(8): 4096–4102.CrossRefGoogle Scholar
  11. 11.
    Jolanta, B. G., Mediyha, S., Susanna, C., Activation of heat-shock response by an adenovirus is essential for virus replication, Nature, 2000, 407: 207–211.CrossRefGoogle Scholar
  12. 12.
    Amici, C., Giorgi, C., Rossi, A., Selective inhibition of virus protein synthesis by prostaglandin A1: A translational block associated withHSP70 synthesis, J. Virol., 1994, 11: 6890–6899.Google Scholar
  13. 13.
    Conti, C., Marco, A., Mastromarino, P., Antiviral effect of hyperthermic treatment in rhinovirus infection, Antimicrob. Agents Chemother., 1999, 43(4): 822–829.Google Scholar
  14. 14.
    He, J. G., The influences of environments on WSSV epidemic and the strategy of controlling, In: Breaking out and controlling of diseases on marine aquaculture species (Xiang, J. H. ed.), Beijing: Ocean Press, 2001, 194–200.Google Scholar
  15. 15.
    Sambrook, J., Russell, D., Molecular Cloning, A Laboratory Manual, 3rd ed, New York: Cold Spring Harbor Laboratory Press, 2001, 979.Google Scholar
  16. 16.
    Chomczynski, P., Sacchi, N., Single step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction, Anal Biochem., 1987, 162: 156–159.CrossRefGoogle Scholar
  17. 17.
    Xiang J. H., Wang B., Liu B., Over 10000 expressed sequence tags fromFenneropenaeus chinesis, Book of Abstracts, World Aquaculture Society, 2002, 837.Google Scholar
  18. 18.
    Shen, Y. Q., Xiang, J. H., Wang, B. et al., 2004. Discovery of immune related factors inFenneropenaeus chinensis by annotation of ESTs, Prog. Natl. Sci., 14(1): 47–54.CrossRefGoogle Scholar
  19. 19.
    Demand, J., Luders, J., Hohfeld, J., The carboxy-terminal domain of HSC70 provides binding sites for a distinct set of chaperone cofactors, Mol. Cell Biol., 1998, 18: 2023–2028.Google Scholar
  20. 20.
    Freeman, B. C., Myers, P. M., Schumacher, R., Identification of a regulatory motif in HSP70 that affects ATPase activity, substrate binding and interaction with HDJ-1, EMBO J., 1995, 14: 2281–2292.Google Scholar
  21. 21.
    Vayssier, M., Leguerhier, F., Fabien, J. F., Cloning and analysis of a Trichinella briotovi gene encoding a cytoplasmic heat shock protein of 72 kD, Parasitology, 1999, 119: 81–93.CrossRefGoogle Scholar
  22. 22.
    Kuwabara, K., Matsumoto, M., Ikeda, J., Purification and characterization of a novel stress protein, the 150-kDa oxygen-regulated protein (ORP150), from cultured rat astrocytes and its expression in ischemic mouse brain, J Biol Chem., 1996, 271: 5025–5032.CrossRefGoogle Scholar
  23. 23.
    Lee Yoon, D., Easton, D., Murawski, M., Identification of a major subfamily of large hsp70-like proteins through the cloning of the mammalian 110-kDa heat shock protein, J. Biol. Chem., 1995, 270: 15725–15733.CrossRefGoogle Scholar
  24. 24.
    Schmid, D., Baici, A., Gehring, H., Kinetics of molecular chaperone action. Science, 1994, 263: 971–973.CrossRefGoogle Scholar
  25. 25.
    Craven, R. A., Tyson, J. R., Stirling, C. J., A novel subfamily of HSP70s in the endoplasmic reticulum, Trends Cell Biol., 1997, 7: 277–282.CrossRefGoogle Scholar
  26. 26.
    Hung, J. J., Cheng, T. J., Chang, M. D., Involvement of heat shock elements and basal transcription elements in the differential induction of the 70-kDa heat shock protein and its cognate by cadmium chloride in 9L rat brain tumor cells, J Cell Biochem., 1998, 71: 21–35.CrossRefGoogle Scholar
  27. 27.
    Kaneko, M., Abe, K., Kogure, K., Saito, H., Correlation between electroconvulsive seizure and HSC70 mRNA induction in mice brain, Neurosci Lett., 1993, 157: 195–198.CrossRefGoogle Scholar
  28. 28.
    Santacruz, H., Vriz, S., Angelier, N., Molecular characterization of a heat shock cognate cDNA of zebrafish, hsc70, and developmental expression of the corresponding transcripts, Dev. Genet., 1997, 21: 223–233.CrossRefGoogle Scholar
  29. 29.
    Boone, A. N., Vijayan, M. M., Constitutive heat shock protein 70 (HSC70) expression in rainbow trout hepatocytes: Effect of heat shock and heavy metal exposure, Comp. Biochem. Physiol. C., 2002, 132: 223–233.Google Scholar
  30. 30.
    Yost, H. J., Lindquist, S., RNA splicing is interrupted by heat shock and is rescued by heat shock protein synthesis, Cell, 1986, 25: 185–193.CrossRefGoogle Scholar
  31. 31.
    Yost, H. J., Lindquist, S., Heat shock proteins affect RNA processing during the heat shock response ofSaccharomyces cerevisiae, Mol. Cell Biol., 1991, 11: 1062–1068.Google Scholar

Copyright information

© Science in China Press 2004

Authors and Affiliations

  • Chuanzhen Jiao
    • 1
    • 2
    • 3
  • Zaizhao Wang
    • 1
  • Fuhua Li
    • 1
  • Chengsong Zhang
    • 1
  • Jianhai Xiang
    • 1
  1. 1.Institute of OceanologyChinese Academy of SciencesQingdaoChina
  2. 2.Department of BiologyDezhou CollegeDezhouChina
  3. 3.Graduate School of Chinese Academy of SciencesBeijingChina

Personalised recommendations