Volume 35 Issue 5
Sep.  2014
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Kou PENG, Cheng-Yuan WANG, Jun-Hua WANG, Jun-Qing SHENG, Jian-Wu SHI, Jian LI, Yi-Jiang HONG. Molecular cloning, sequence analysis, and cadmium stress-rated expression changes of BTG1 in freshwater pearl mussel (Hyriopsis schlegelii). Zoological Research, 2014, 35(5): 389-397. doi: 10.13918/j.issn.2095-8137.2014.5.389
Citation: Kou PENG, Cheng-Yuan WANG, Jun-Hua WANG, Jun-Qing SHENG, Jian-Wu SHI, Jian LI, Yi-Jiang HONG. Molecular cloning, sequence analysis, and cadmium stress-rated expression changes of BTG1 in freshwater pearl mussel (Hyriopsis schlegelii). Zoological Research, 2014, 35(5): 389-397. doi: 10.13918/j.issn.2095-8137.2014.5.389

Molecular cloning, sequence analysis, and cadmium stress-rated expression changes of BTG1 in freshwater pearl mussel (Hyriopsis schlegelii)

doi: 10.13918/j.issn.2095-8137.2014.5.389
Funds:  This work was supported by the Key Scientific and Technological Programme of Jiangxi Province, China (20121BBF60036); the Special Fund for Agro-scientific Research in the Public Interest, State Agriculture Ministry of China (200903028); the Science and Technology Landing Project of Jiangxi Province, China (KJLD12001); the Youth Fund of the Education Department of Jiangxi Province, China (GJJ14219) and the National Natural Science Foundation of China (31160534)
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  • Corresponding author: Yi-Jiang HONG
  • Received Date: 2013-12-12
  • Rev Recd Date: 2014-08-04
  • Publish Date: 2014-09-08
  • The B cells translocation gene 1 (BTG1) is a member of the BTG/TOB family of anti-proliferative genes, which have recently emerged as important regulators of cell growth and differentiation among vertebrates. Here, for the first time we cloned the full-length cDNA sequence of Hyriopsis schlegelii (Hs-BTG1), an economically important freshwater shellfish and potential indicator of environmental heavy metal pollution, for the first time. Using rapid amplification of cDNA ends (RACE) together with splicing the EST sequence from a haemocyte cDNA library, we found that Hs-BTG1 contains a 525 bp open reading frame (ORF) encoding a 174 amino-acid polypeptide, a 306 bp 5' untranslated region (5' UTR), and a 571 bp 3' UTR with a Poly(A) tail as well as a transcription termination signal (AATAAA). Homologue searching against GenBank revealed that Hs-BTG1 was closest to Crassostrea gigas BTG1, sharing 50.57% of protein identities. Hs-BTG1 also shares some typical features of the BTG/TOB family, possessing two well-conserved A and B boxes. Clustering analysis of Hs-BTG1 and other known BTGs showed that Hs-BTG1 was also closely related to BTG1 of C. gigas from the invertebrate BTG1 clade. Function prediction via homology modeling showed that both Hs-BTG1 and C. gigas BTG1 share a similar three-dimensional structure with Homo sapiens BTG1. Tissue-specific expression analysis of the Hs-BTG1 via real-time PCR showed that the transcripts were constitutively expressed, with the highest levels in the hepatopancreas and gills, and the lowest in both haemocyte and muscle tissue. Expression levels of Hs-BTG1 in hepatopancreas (2.03-fold), mantle (2.07-fold), kidney (2.2-fold) and haemocyte (2.5-fold) were enhanced by cadmium (Cd2+) stress, suggesting that Hs-BTG1 may have played a significant role in H. schlegelii adaptation to adverse environmental conditions.
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  • [1] Busson M, Carazo A, Seyer P, Grandemange S, Casas F, Pessemesse L, Rouault JP, Wrutniak-Cabello C, Cabello G. 2005. Coactivation of nuclear receptors and myogenic factors induces the major BTG1 influence on muscle differentiation. Oncogene, 24(10): 1698-710.
    [2] Corjay MH, Kearney MA, Munzer DA, Diamond SM, Stoltenborg JK. 1998. Antiproliferative gene BTG1 is highly expressed in apoptotic cells in macrophage-rich areas of advanced lesions in Watanabe heritable hyperlipidemic rabbit and human. Laboratory Investigation, 78: 847-858.
    [3] Duan YF, Liu P, Li JT, Li J, Gao BQ, Chen P. 2013. Cloning and expression analysis of Cathepsin L cDNA of Exopalaemon carinicauda. Zoological Research, 34(1): 39-46.(in Chinese)
    [4] Fu YJ, Huang FG, Yuan T, Gu JR, Luo GQ, Xu H. 2012. Molecular cloning, characterization and expression analysis of B cell translocation gene 1 in grass carp Ctenopharyngodon idella. Journal of Fish Biology, 80(3): 669-678.
    [5] Feng Z, Shen H, Du ZQ, Zhu MJ, Fan B, Rothschild MF, Zhao SH, Li CC. 2011. BTG1 as a new candidate gene for muscle growth in pigs: Cloning, expression and association analysis. Journal of Animal Science and Biotechnology, 2(3): 121-130.
    [6] Guehenneux F, Duret L, Callanan MB, Bouhas R, Hayette S, Berthet C, Samarut C, Rimokh R, Birot AM, Wang Q, Magaud JP, Rouault JP. 1997. Cloning of the mouse BTG3 gene and definition of a new gene family (the BTG family) involved in the negative control of the cell cycle. Leukemia, 11(3): 370-375.
    [7] He SH, Peng K, Hong YJ, Wang JH, Sheng JQ, Gu Q. 2013. Molecular properties and immune defense of two ferritin subunits from freshwater pearl mussel, Hyriopsis schlegelii. Fish & Shellfish Immunology, 34(3): 865-874.
    [8] Hata K, Nishijima K, Mizuguchi J. 2007. Role for Btg1 and Btg2 in growth arrest of WEHI-231 cells through arginine methylation following membrane immunoglobulin engagement. Experimental Cell Research, 313(11): 2356-2366.
    [9] Iwai K, Hirata K, Ishida T, Takeuchi S, Hirase T, Rikitake Y, Kojima Y, Inoue N, Kawashima S, Yokoyama M. 2004. An anti-proliferative gene BTG1 regulates angiogenesis in vitro. Biochemical and Biophysical Research Communications, 316(3): 628-635.
    [10] Kim JH, Rhee JS, Dahms HU, Lee YM, Han KN, Lee JS. 2012. The yellow catfish, Pelteobagrus fulvidraco (Siluriformes) metallothionein cDNA: molecular cloning and transcript expression level in response to exposure to the heavy metals Cd, Cu, and Zn. Fish Physiology and Biochemistry, 38(5): 1331-1342.
    [11] Lee H, Cha S, Lee MS, Cho GJ, Choi WS, Suk K. 2003. Role of antiproliferative B cell translocation gene-1 as an apoptotic sensitizer in activation-induced cell death of brain microglia. Journal of Immunology, 171(11): 5802-5811.
    [12] Lu CH, Xie WZ, Zhou GJ. 1998. Studies on Crassostrea rivularis as a biological indicator of cadmium pollution. Journal of Fishery Science of China, 5(2): 79-83.(in Chinese)
    [13] Mauxion F, Chen CY, Séraphin B, Shyu AB. 2009. BTG/TOB factors impact deadenylases. Trends in Biochemical Sciences, 34(12): 640-647.
    [14] Melamed J, Kernizan S, Walden PD. 2002. Expression of B-cell translocation gene 2 protein in normal human tissues. Tissue Cell, 34(1): 28-32.
    [15] Matsuda S, Rouault J, Magaud J, Berthet C. 2001.In search of a function for the TIS21/PC3/BTG1/TOB family. FEBS Letters, 497(2-3): 67-72.
    [16] Peng K, Wang JH, Sheng JQ, Zeng LG, Hong YJ. 2012a. Molecular characterization and immune analysis of a defensin from freshwater pearl mussel, Hyriopsis schlegelii. Aquaculture, 334-337: 45-50.
    [17] Peng K, Wang JH, Liu TT, Sheng JQ, Shi JW, Shao P, He SH, Hong YJ. 2012b. Expression analysis and immune response of the cathepsin L from freshwater pearl mussel, Hypriopsis schlegelii Acta Hydrobiologica Sinca, 36(6): 1128-1134.(in Chinese)
    [18] Rouault JP, Rimokh R, Tessa C, Paranhos G, Ffrench M, Duret L, Garoccio M, Germain D, Samarut J, Magaud JP. 1992. BTG1, a member of a new family of antiproliferative genes. EMBO Journal, 11(4): 1663-1670.
    [19] Rodier A, Rochard P, Berthet C, Rouault JP, Casas F, Daury L, Busson M, Magaud JP, Wrutniak-Cabello C, Cabello G. 2001. Identification of functional domains involved in BTG1 cell localization. Oncogene, 20(21): 2691-2703.
    [20] Sakaguchi T, Kuroiwa A, Takeda H. 2001. Expression of zebrafish btg-b, an anti-proliferative cofactor, during early embryogenesis. Mechanisms of Development, 104(1-2): 113-115.
    [21] Saka Y, Tada M, Smith JC. 2000. A screen for targets of the Xenopus T-box gene Xbra. Mechanisms of Development, 93(1-2): 27-39.
    [22] Wang ZS, Qi ZT, Tian JY, Qiu M, Zhao WH, Wang AM, Huang JT, Guo XJ. 2011. Cloning of hemoglobin-α1 from half-smooth tongue sole (Cynoglossus semilaevis) and its expression under short-term hypoxia. Zoological Research, 32(6): 641-646.
    [23] Wessely O, Kim JI, Tran U, Fuentealba L, De Robertis EM. 2005. xBtg-x regulates Wnt/beta-Catenin signaling during early Xenopus development. Developmental Biology, 283(1): 17-28.
    [24] Winkler GS. 2010. The mammalian anti-proliferative BTG/Tob protein family. Journal of Cellular Physiology, 222(1): 66-72.
    [25] Xie K, Xu L, Sheng JQ, Zeng LG, Wang JH, Hong YJ. 2011. The full-length cDNA library of hemocyte induced by Aeromonas hydrophila and molecular characteristics of Cyclophilin A from Hyriopsis schlegelii. Acta Hydrobiologica Sinica, 35(5): 783-789.(in Chinese)
    [26] Yang X, Morita M, Wang H, Suzuki T, Yang W, Luo Y, Zhao C, Yu Y, Bartlam M, Yamamoto T, Rao Z. 2008. Crystal structures of human BTG2 and mouse TIS21 involved in suppression of CAF1 deadenylase activity. Nucleic Acids Research, 36(21): 6872-6881.
    [27] Zhang J, Du LX, Li HB, Wei CH. 2009. Semi-quantitative RT-PCR and bioinformatics analysis of sheep BTG1 gene. Chinese Journal of Biochemistry and Molecular Biology, 25(10): 911-918.(in Chinese)
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Molecular cloning, sequence analysis, and cadmium stress-rated expression changes of BTG1 in freshwater pearl mussel (Hyriopsis schlegelii)

doi: 10.13918/j.issn.2095-8137.2014.5.389
Funds:  This work was supported by the Key Scientific and Technological Programme of Jiangxi Province, China (20121BBF60036); the Special Fund for Agro-scientific Research in the Public Interest, State Agriculture Ministry of China (200903028); the Science and Technology Landing Project of Jiangxi Province, China (KJLD12001); the Youth Fund of the Education Department of Jiangxi Province, China (GJJ14219) and the National Natural Science Foundation of China (31160534)
    Corresponding author: Yi-Jiang HONG

Abstract: The B cells translocation gene 1 (BTG1) is a member of the BTG/TOB family of anti-proliferative genes, which have recently emerged as important regulators of cell growth and differentiation among vertebrates. Here, for the first time we cloned the full-length cDNA sequence of Hyriopsis schlegelii (Hs-BTG1), an economically important freshwater shellfish and potential indicator of environmental heavy metal pollution, for the first time. Using rapid amplification of cDNA ends (RACE) together with splicing the EST sequence from a haemocyte cDNA library, we found that Hs-BTG1 contains a 525 bp open reading frame (ORF) encoding a 174 amino-acid polypeptide, a 306 bp 5' untranslated region (5' UTR), and a 571 bp 3' UTR with a Poly(A) tail as well as a transcription termination signal (AATAAA). Homologue searching against GenBank revealed that Hs-BTG1 was closest to Crassostrea gigas BTG1, sharing 50.57% of protein identities. Hs-BTG1 also shares some typical features of the BTG/TOB family, possessing two well-conserved A and B boxes. Clustering analysis of Hs-BTG1 and other known BTGs showed that Hs-BTG1 was also closely related to BTG1 of C. gigas from the invertebrate BTG1 clade. Function prediction via homology modeling showed that both Hs-BTG1 and C. gigas BTG1 share a similar three-dimensional structure with Homo sapiens BTG1. Tissue-specific expression analysis of the Hs-BTG1 via real-time PCR showed that the transcripts were constitutively expressed, with the highest levels in the hepatopancreas and gills, and the lowest in both haemocyte and muscle tissue. Expression levels of Hs-BTG1 in hepatopancreas (2.03-fold), mantle (2.07-fold), kidney (2.2-fold) and haemocyte (2.5-fold) were enhanced by cadmium (Cd2+) stress, suggesting that Hs-BTG1 may have played a significant role in H. schlegelii adaptation to adverse environmental conditions.

Kou PENG, Cheng-Yuan WANG, Jun-Hua WANG, Jun-Qing SHENG, Jian-Wu SHI, Jian LI, Yi-Jiang HONG. Molecular cloning, sequence analysis, and cadmium stress-rated expression changes of BTG1 in freshwater pearl mussel (Hyriopsis schlegelii). Zoological Research, 2014, 35(5): 389-397. doi: 10.13918/j.issn.2095-8137.2014.5.389
Citation: Kou PENG, Cheng-Yuan WANG, Jun-Hua WANG, Jun-Qing SHENG, Jian-Wu SHI, Jian LI, Yi-Jiang HONG. Molecular cloning, sequence analysis, and cadmium stress-rated expression changes of BTG1 in freshwater pearl mussel (Hyriopsis schlegelii). Zoological Research, 2014, 35(5): 389-397. doi: 10.13918/j.issn.2095-8137.2014.5.389
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