Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (17): 3592-3601.doi: 10.3864/j.issn.0578-1752.2012.17.016

• ANIMAL SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

Cloning and Sequence Analysis of JAK/STAT Pathway Related Major Genes of Silkworm (Bombyx mori)

 PENG  Jing-Kun, CAO  Guang-Li, QIAN  Ying, ZHANG  Xiao-Li, MA  Huan-Yan, HE  Lei, XUE  Ren-Yu, GONG  Cheng-Liang   

  1. 1.苏州大学基础医学与生物科学学院,江苏苏州 215123
  • Received:2011-09-23 Online:2012-09-01 Published:2012-03-26

Abstract: 【Objective】The objective of this study is to identify the elements of JAK/STAT pathway of silkworm, Bombyx mori and determine the function of JAK/STAT pathway related major genes in silkworm. 【Method】JAK/STAT pathway related major genes in silkworm were obtained through electronic cloning and conventional RT-PCR. Domain analysis on the related protein were conducted by using bio-softwares online, and the expression profile of these genes in diverse tissues were analysed through microarray database or qPCR.【Result】The silkworm JAK/STAT pathway related major genes including BmSTAT, BmHOP, BmSOCS2, BmSOCS5A, BmSOCS5B, BmSOCS6, BmDRK, Bmken, BmPIAS1, BmPIAS2 and BmPI3K were cloned, respectively, but the homologues of Drosophila Upd1, Upd2 and Upd3 were not cloned. Sequence analysis showed that BmHOP contained CRD_FZ, Kringle domains, transmembrane region and Pkc superfamily protein kinase catalysis domain TyrKc. BmSTAT had two isoforms, which were consisted of SH2, STAT_bind, STAT_int and STAT_alpha domains. Both BmSOCS2A and BmSOCS6A had a conserved SH2 domain and a SOCS box in cloned three genes of BmSOCS family. BmDRK was consisted of SH3 and SH2 domains that connected in series to form SH3-SH2-SH3. BmKen contained BTB and zinc finger domains. BmPIAS1 and BmPIAS2 contained LCR domains. BmPI3K had two conserved SH2 domains.【Conclusion】The JAK/STAT pathway related major genes were cloned successfully in silkworm. The analysis of conserved structural domains suggests that BmHOP is one of the JAK kinases, which phosphorylates the BmSTAT transcription factor and leads to activation of target genes. BmSOCS members, Bmken, BmPIAS1 and BmPIAS2 are negative feedback regulators in the JAK/STAT pathway.

Key words: Bombyx mori, JAK/STAT signaling pathway, gene cloning, sequence analysis

[1]Brivanlou A H, Darnell Jr J E. Signal transduction and the control of gene expression. Science, 2002, 295(5556): 813-818.

[2]Hombria J C G, Brown S. The fertile field of Drosophila JAK/STAT signalling. Current Biology, 2002, 12(16): 569-575.

[3]Wawersik M, Milutinovich A, Casper A L, Matunis E, Williams B, Van Doren M. Somatic control of germline sexual development is mediated by the JAK/STAT pathway. Nature, 2005, 436(7050): 563-567.

[4]Harrison D A, McCoon P E, Binari R, Gilman M, Perrimon N. Drosophila unpaired encodes a secreted protein that activates the JAK signaling pathway. Genes & Development, 1998, 12(20): 3252-3263.

[5]Aringer M, Cheng A, Nelson J W, Chen M, Sudarshan C, Zhou Y J, O'Shea J J. Janus kinases and their role in growth and disease. Life Science, 1999, 64(24): 2173-2186.

[6]Hansen J A, Lindberg K, Hilton D J, Nielsen J H, Billestrup N. Mechanism of inhibition of growth hormone receptor signaling by suppressor of cytokine signaling proteins. Molecular Endocrinology, 1999, 13(11): 1832-1843.

[7]Ram P A, Waxman D J. SOCS/CIS protein inhibition of growth hormone-stimulated STAT5 signaling by multiple mechanisms. The Journal of Biological Chemistry, 1999, 274(50): 35553-35561.

[8]Cooney R N. Suppressors of cytokine signaling (SOCS): inhibitors of the JAK/STAT pathway. Shock, 2002, 17(2): 83-90.

[9]Wormald S, Hilton D J. Inhibitors of cytokine signal transduction. The Journal of Biological Chemistry, 2004, 279(2): 821-824.

[10]Banerjee U, Renfranz P J, Pollock J A, Benzer S. Molecular characterization and expression of sevenless, a gene involved in neuronal pattern formation in the Drosophila eye. Cell, 1987, 49(2): 281-291.

[11]Hafen E, Basler K, Edstroem J E, Rubin G M. Sevenless, a cell-specific homeotic gene of Drosophila, encodes a putative transmembrane receptor with a tyrosine kinase domain. Science, 1987, 236(4797): 55-63.

[12]Tomlinson A, Ready D F. Sevenless: a cell-specific homeotic mutation of the Drosophila eye. Science, 1986, 231(4736): 400-402.

[13]Almudi I, Corominas M, Serras F. Competition between SOCS36E and Drk modulates Sevenless receptor tyrosine kinase activity. Journal of Cell Science, 2010, 123(22): 3857-3862.

[14]Arbouzova N I, Bach E A, Zeidler M P. Ken & barbie selectively regulates the expression of a subset of Jak/STAT pathway target genes. Current Biology, 2006, 16(1): 80-88.

[15]Chung C D, Liao J, Liu B, Rao X, Jay P, Berta P, Shuai K. Specific inhibition of Stat3 signal transduction by PIAS3. Science, 1997, 278(5344): 1803-1805.

[16]Betz A, Lampen N, Martinek S, Young M W, Darnell Jr J E. A Drosophila PIAS homologue negatively regulates stat92E. Proceedings of Natural Academic Sciences of the United Sataes of America, 2001, 98(17): 9563-9568.

[17]牛国梁, 张树友. PI3K/Akt信号传导通路与肿瘤. 现代生物医学进展, 2010, 10(20): 3994-3996.

Niu G L, Zhang S Y. PI3K/Akt signaling pathways and tumors. Progress in Modern Biomedicine, 2010, 10(20): 3994-3996. (in Chinese)

[18]Xia Q, Zhou Z, Lu C, Cheng D, Dai F, Li B, Zhao P, Zha X, Cheng T, Chai C, Pan G, Xu J, Liu C, Lin Y, Qian J, Hou Y, Wu Z, Li G, Pan M, Li C, Shen Y, Lan X, Yuan L, Li T, Xu H, Yang G, Wan Y, Zhu Y, Yu M, Shen W, Wu D, Xiang Z, Yu J, Wang J, Li R, Shi J, Li H, Li G, Su J, Wang X, Li G, Zhang Z, Wu Q, Li J, Zhang Q, Wei N, Xu J, Sun H, Dong L, Liu D, Zhao S, Zhao X, Meng Q, Lan F, Huang X, Li Y, Fang L, Li C, Li D, Sun Y, Zhang Z, Yang Z, Huang Y, Xi Y, Qi Q, He D, Huang H, Zhang X, Wang Z, Li W, Cao Y, Yu Y, Yu H, Li J, Ye J, Chen H, Zhou Y, Liu B, Wang J, Ye J, Ji H, Li S, Ni P, Zhang J, Zhang Y, Zheng H, Mao B, Wang W, Ye C, Li S, Wang J, Wong G K S, Yang H. A draft sequence for the genome of the domesticated silkworm (Bombyx mori). Science, 2004, 306(5703): 1937-1940.

[19]Rawlings J S, Rosler K M, Harrison D A. The JAK/STAT signaling pathway. Journal of Cell Science, 2004, 117(8): 1281-1283.

[20]Cheng T C, Xia Q Y, Xu P Z, Tan X, Fang T, Xiang Z H. Identification and comparative analysis of immune-related genes and signaling pathways in the silkworm, Bombyx mori. Acta Entomologica Sinica, 2009, 52(3): 235-245.

[21]Hilton D J. Negative regulators of cytokine signal transduction. Cellular and Molecular Life Sciences, 1999, 55(12): 1568-1577.

[22]Hou S X, Zheng Z, Chen X, Perrimon N. The JAK/STAT pathway in model organisms: emerging roles in cell movement. Developmental Cell, 2002, 3(6): 765-778.

[23]Rawlings J S, Rennebeck G, Harrison S M, Xi R, Harrison D A. Two Drosophila suppressors of cytokine signaling (SOCS) differentially regulate JAK and EGFR pathway activities. BMC Cell Biology, 2004, 5(1): 38.

[24]Arbouzova N I, Zeidler M P. JAK/STAT signalling in Drosophila: insights into conserved regulatory and cellular functions. Development, 2006, 133(14): 2605-2616.

[25]Bina S, Wright V M, Fisher K H, Milo M, Zeidler M P. Transcriptional targets of Drosophila JAK/STAT pathway signalling as effectors of haematopoietic tumour formation. EMBO Reports, 2010, 11(3): 201-207.

[26]Ihle J N, Kerr I M. Jaks and Stats in signaling by the cytokine receptor superfamily. Trends in Genetics, 1995, 11(2): 69-74.

[27]Imada K, Leonard W J. The Jak-STAT pathway. Molecular Immunology, 2000, 37(1/2): 1-11.
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