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ACOMS+ 및 학술지 리포지터리 설명회

  • 한국과학기술정보연구원(KISTI) 서울분원 대회의실(별관 3층)
  • 2024년 07월 03일(수) 13:30
 

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북방전복 (Haliotis discus hannai) 의 열 충격 결합 단백질1 (HspBP1, Heat shock binding protein1) 유전자 동정 및 발현분석

Molecular cloning, characterization and expression analysis of heat shock binding protein1 from the pacific abalone (Haliotis discus hannai)

Abstract

Abalone is economically important marine resources for fisheries and aquaculture industries worldwide. The climate changes cause the mass mortality of farmed abalone and have led to the economic loss of fisheries and aquaculture industries in Korea. In order to find genes which are involved in physiological response to temperature stress, we have performed sequencing of transcriptome after thermal stimulation. The heat-shock response is elicited by exposure of cells to thermal and chemical stress and induces activation of Hsp70. Heat shock binding protein 1 (HspBP1) was originally identified as a protein that interacts with Hsp70 and inhibits its chaperone. In this study, molecular characterization and expression of HspBP1, named abHspBP1, were studied to understand the mechanism of heat shock response of the pacific abalone (Haliotis discus hannai) under thermal stresses. The full-length cDNA of abHspBP1 was of 981 bp, encoding a protein of 326 aa. Multiple alignments and phylogenetic analysis with the deduced amino acid of abHspBP1 showed homology with Mizuhopectenyessoensis HspBP1. Reverse transcription PCR analysis indicated that the abHspBP1 mRNA in the hemocyte and gill was drastically increased at 30°C. The expression of Hsp70 in the hemocytes and gills was drastically increased at 30°C. Our present study for the first time reported the identification, molecular cloning, characterization and tissue expression analysis of HspBP1 in Haliotis discus hannai. The results of this study might provide new insights into the regulation of heat shock response in this species.

keywords
Haliotis discus hannai, heat shock binding protein 1, heat stress, mRNA expression, cDNA cloning

참고문헌

1.

Livak, K.J. and Schmittgen, T.D. (2007) Analysis ofrelative gene expression data using real-time quantitative PCR and the <TEX>$2^{-{\Delta}{\Delta}CT}$</TEX> method. Methods, 25: 402-408.

2.

Alberti, S., Bohse, K., Arndt, V., Schmitz, A., and Hohfeld, J. (2004) The cochaperone HspBP1 inhibits the CHIP ubiquitin ligase and stimulates the maturation of the cystic fibrosis transmembrane conductance regulator. Molecular biology of the cell, 15(9): 4003-4010.

3.

Ballinger, C.A., Connell, P., Wu, Y., Hu, Z., Thompson, L.J., Yin, L.Y., and Patterson, C. (1999) Identification of CHIP, a novel tetratricopeptide repeat-containing protein that interacts with heat shock proteins and negatively regulates chaperone functions. Mollecular and Cellular Biology, 19(6): 4535-4545.

4.

Bardelli, A., Longati, P., Albero, D., Goruppi, S., Schneider, C., Ponzetto, C., and Comoglio, P.M. (1996) HGF receptor associates with the anti-apoptotic protein BAG-1 and prevents cell death. EMBO Journal, 15(22): 6205-621

5.

Brehme, M., Voisine, C., Rolland, T., Wachi, S., Soper, J.H., Zhu, Y., Orton, K., Villella, A., Garza, D., Vidal, M., Ge, H. and Morimoto, R.I. (2014) A chaperome subnetwork safeguards proteostasis in aging and neurodegenerative disease. Cell Reports, 9(3): 1135-1150.

6.

Bukau, B. and Horwich, A.L. (1988) The Hsp70 and Hsp60 chaperone machines. Cell, 92(3): 351-366.

7.

DeLuca-Flaherty C1, McKay DB, Parham P, Hill BL. (1990) Uncoating protein (hsc70) binds a conformationally labile domain of clathrin light chain LCa to stimulate ATP hydrolysis. Cell, 62(5): 875-887.

8.

Bukau, B., Deuerling, E., Pfund, C., and Craig, E.A. (2000) Getting Newly Synthesized Proteins into Shape. Cell, 101(2): 119-122.

9.

Chung, K.T., Shen, Y., and Hendershot, L.M. (2002) BAP, a mammalian BiP-associated protein, is a nucleotide exchange factor that regulates the ATPase activity of BiP. Journal of biological chemistry, 277(49): 47557-47563

10.

Frydman, J. (2001) Folding of newly translated proteins in vivo: the role of molecular chaperones. Annual Review of Biochemistry, 70: 603-647.

11.

Graner, M.W., Raynes, D.A., Bigner, D.D., and Guerriero, V. (2009) Heat shock protein 70-binding protein 1 is highly expressed in high-grade gliomas, interacts with multiple heat shock protein 70 family members, and specifically binds brain tumor cell surfaces. cancer science, 100(10): 1870-1879.

12.

Hartl, F.U. and Hayer-Hartl, M. (2002) Molecular chaperones in the cytosol: from nascent chain to folded protein. Science, 295(5561): 1852-1858.

13.

Hohfeld, J., Minami, Y., and Hartl, F.U. (1995) Hip, a novel cochaperone involved in the eukaryotic Hsc70/Hsp40 reaction cycle. Cell, 83(4): 589-598.

14.

Kabani, M., McLellan, C., Raynes, D.A., Guerriero, V., and Brodsky, J.L. (2002) HspBP1, a homologue of the yeast Fes1 and Sls1 proteins, is an Hsc70 nucleotide exchange factor. FEBS Letters, 531(2): 339-342.

15.

Kabani, M., Beckerich, J.M., and Gaillardin, C. (2000) Sls1p stimulates Sec63p-mediated activation of Kar2p in a conformation-dependent manner in the yeast endoplasmic reticulum. Mollecular and Cellular Biology, 20(18): 6923-6934.

16.

Karlin, S. and Altschul, S.F. (1990) Methods for assessing the statistical significance of molecular sequence features by using general scoring schemes. Proceedings of the National Academy of Sciences of the United States of America, 87(6): 2264-2268.

17.

Karlin, S. and Altschul, S.F. (1993) Applications and statistics for multiple high-scoring segments in molecular sequences. Proceedings of the National Academy of Sciences of the United States of America, 90(12): 5873-5877.

18.

Lindquist, S. and Craig, E.A. (1988) The heat-shock proteins. Annual Review of Genetics, 22: 631-677.

19.

Mayer, M.P. and Bukau, B. (2005) Hsp70 chaperones: cellular functions and molecular mechanism. Cellular and Molecular Life Science, 62(6): 670-684.

20.

McLellan, C.A., Raynes, D.A., and Guerriero, V. (2004) HspBP1, an Hsp70 cochaperone, has two structural domains and is capable of altering the conformation of the Hsp70 ATPase domain. Journal of biological chemistry, 278(21): 19017-19022.

21.

Prapapanich, V., Chen, S., Nair, S.C., Rimerman, R.A., and Smith, D.F. (1996) Molecular cloning of human p48, a transient component of progesterone receptor complexes and an Hsp70-binding protein. Molecular Endocrinology, 10(4): 420-431.

22.

Raynes, D.A. and Guerriero, V.Jr. (1998) Inhibition of Hsp70 ATPase activity and protein renaturation by a novel Hsp70-binding protein. Journal of biological chemistry, 273(49): 32883-32888.

23.

Raynes, D.A., Thomson, C.A., Stroster, J., Newton, T., Cuneo, P., and Guerriero, V. (2006) Human serum contains detectable levels of the Hsp70 cochaperone HspBP1 and antibodies bound to HspBP1. Journal of Immunoassay & Immunochemistry, 27(3): 251-264.

24.

Shomura, Y., Dragovic, Z., Chang, H.C., Tzvetkov, N., Young, J.C., Brodsky, J.L., Guerriero, V., Hartl, F.U., and Bracher, A. (2005) Regulation of Hsp70 function by HspBP1: structural analysis reveals an alternate mechanism for Hsp70 nucleotide exchange. Molecular Cell, 17(3): 367-379.

25.

Tanimura, S., Hirano, A.I., Hashizume, J., Yasunaga, M., Kawabata, T., Ozaki, K., and Kohno, M. (2007) Anticancer drugs up-regulate HspBP1 and thereby antagonize the prosurvival function of Hsp70 in tumor cells. Journal of biological chemistry, 282(49): 35430-35439.

26.

Wu, S.J., Liu, F.H., Hu, S.M., and Wang, C. (2001) Different combinations of the heat-shock cognate protein 70 (hsc70) C-terminal functional groups are utilized to interact with distinct tetratricopeptide repeat-containing proteins. Biochemical Journal, 359(Pt 2): 419-426.

27.

Yang, X., Chernenko, G., Hao, Y., Ding, Z., Pater, M.M., Pater, A., and Tang, S.C. (1998) Human BAG-1/RAP46 protein is generated as four isoforms by alternative translation initiation and over expressed in cancer cells. Oncogene, 17(8): 981-989.

28.

Young, J.C., Agashe, V.R., Siegers, K., and Hartl, F.U. (2004) Pathways of chaperone-mediated protein folding in the cytosol. Nature Reviews Molecular Cell Biology, 5(10): 781-791.

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