家蚕JAK/STAT信号通路相关基因克隆与分析

doi:10.3864/j.issn.0578-1752.2012.17.016

摘要/Abstract

摘要： 【目的】鉴定家蚕JAK/STAT信号通路组成原件，并预测家蚕JAK/STAT信号通路主要相关基因的功能。【方法】在电子克隆的基础上，通过RT-PCR获得家蚕JAK/STAT信号通路的主要相关基因；利用在线软件对相关蛋白进行结构域分析；基于microarray数据库或qPCR分析各基因的组织表达特征。【结果】克隆了家蚕JAK/STAT信号通路主要相关基因，包括BmSTAT、BmHOP、BmSOCS2、BmSOCS5A、BmSOCS5B、BmSOCS6、BmDRK、Bmken、BmPIAS1、BmPIAS2以及BmPI3K，但没有能克隆到果蝇Upd1、Upd2和Upd3的同源体基因。序列分析显示BmHOP具CRD_FZ、Kringle、转膜区域和PKc超家族蛋白激酶催化结构域TyrKc；BmSTAT有2种亚型，均具SH2、STAT_bind、STAT_int和STAT_alpha结构域；在克隆的BmSOCS家族的3个基因中，BmSOCS2A和BmSOCS6A具典型的SH2和SOCS结构域；BmDRK由SH3和SH2两种结构域形成了SH3-SH2-SH3串联结构；BmKen存在BTB和锌指结构域，BmPIAS1和BmPIAS2具有LCR结构域，BmPI3K具有2个SH2结构域。【结论】成功克隆了家蚕JAK/STAT信号通路主要相关基因。相关蛋白的保守结构域分析显示，在家蚕JAK/STAT信号通路中，BmHOP是JAK激酶的一种，可以磷酸化BmSTAT转录因子，激活靶基因转录；BmSOCS、Bmken、BmPIAS1以及BmPIAS2对JAK/STAT通路起负反馈作用。

关键词: 家蚕, JAK/STAT通路, 基因克隆, 序列分析

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

彭景琨, 曹广力, 钱莹, 张晓丽, 马焕艳, 何蕾, 薛仁宇, 贡成良. 家蚕JAK/STAT信号通路相关基因克隆与分析[J]. 中国农业科学, 2012, 45(17): 3592-3601.

PENG Jing-Kun, CAO Guang-Li, QIAN Ying, ZHANG Xiao-Li, MA Huan-Yan, HE Lei, XUE Ren-Yu, GONG Cheng-Liang. Cloning and Sequence Analysis of JAK/STAT Pathway Related Major Genes of Silkworm (Bombyx mori)[J]. Scientia Agricultura Sinica, 2012, 45(17): 3592-3601.

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参考文献

[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.