文昌鱼AmphiRab23b基因的克隆、进化分析及表达模式
收稿日期: 2009-08-25
修回日期: 2009-09-22
网络出版日期: 2009-12-10
基金资助
国家自然科学基金项目(编号:30570967, 30671072, 30700434)和国家重点基础研究发展规划(973计划)项目(编号:2007CB947100, 2007CB815800)资助
Isolation, phylogenetic analysis and developmental expression part-tern of AmphiRab23b in amphioxus
Received date: 2009-08-25
Revised date: 2009-09-22
Online published: 2009-12-10
Hedgehog信号通路在胚胎发育过程中发挥着重要作用, 同时与多种肿瘤的发生密切相关。Rab23蛋白在Hedgehog信号通路中扮演着十分重要的角色。目前关于文昌鱼Rab23同源基因的研究仅局限于佛罗里达文昌鱼(Branchiostoma floridae)基因组中的注释。文章首次克隆了中国文昌鱼(Branchiostoma belcheri) Rab23b基因 (AmphiRab23b)cDNA全序列 , 对其演译的蛋白序列进行了序列比对、进化树分析以及基因时空表达分析。研究结果显示, 文昌鱼AmphiRab23b基因的 cDNA总长为2 062 bp (包括UTR区), 其中开放阅读框 (Open reading frame, ORF) 714 bp, 编码237个氨基酸; 虽然在进化树中不属于脊椎动物Rab23进化支, 但是AmphiRab23具有保守的Rab23_lke结构域, 暗示该基因在进化过程中可能在功能上是保守的。时空表达的研究结果进一步显示, AmphiRab23b基因在胚胎发育中的神经板和消化道中表达, 与其脊椎动物同源基因的表达模式相似, 这说明该基因可能对文昌鱼神经系统和消化道的发育有重要作用。
关键词: 文昌鱼; AmphiRab23; 进化; 表达
李建伟,林浴霜,陈冬艳,张红卫 . 文昌鱼AmphiRab23b基因的克隆、进化分析及表达模式[J]. 遗传, 2009 , 31(12) : 1233 -1240 . DOI: 10.3724/SP.J.1005.2009.01233
The hedgehog (Hh) pathway plays an important role during the embryonic development and is related to the progression of cancers. Rab23 is a crucial functional molecule in Hh pathway. However, there is no report about amphioxus Rab23 up to now except the annotations of two isoforms in the genome of Florida lancelet (Branchiostoma floridae). Here a 2062 bp full-length cDNA sequence of the Rab23, AmphiRab23b, was isolated from Chinese amphioxus (Branchiostoma belcheri), which included the UTRs and an open reading frame of 714 bp, encoding a protein of 237 amino acids. Phylogenetic analysis suggested that AmphiRab23b falled outside the vertebrate clade. But sequence analysis indicated that this putative AmphiRab23b protein contained a specific Rab23_lke domain, which implied that Rab23 gene was functional conservative during evolution. And its developmental expression pattern showed that AmphiRab23b was expressed in the differentiating neural plate and alimentary canal, as the same as the expression pattern of the homologous vertebrate genes, which suggested that AmphiRab23b may function in the development of nervous system and alimentary canal.
Key words: amphioxus; Amphirab23b; evolution; expression
[1] Novick P, Field C, Schekman R. Identification of 23 com-plementation groups required for post-translational events in the yeast secretory pathway. Cell, 1980, 21(1): 205-215.
[2] Gallwitz D, Donath C, Sander C. A yeast gene encoding a protein homologous to the human c-has/bas proto-oncogene product. Nature, 1983, 306(5944): 704-707.
[3] Touchot N, Chardin P, Tavitian A. Four additional mem-bers of the ras gene superfamily isolated by an oligonu-cleotide strategy: molecular cloning of ypt-related cdnas from a rat brain library. Proc Natl Acad Sci USA, 1987, 84(23): 8210-8214.
[4] Entchev EV, Schwabedissen A, Gonzalez-Gaitan M. Gra-dient formation of the tgf-beta homolog dpp. Cell, 2000, 103(6): 981-991.
[5] Lanzetti L, Rybin V, Malabarba MG, Christoforidis S, Scita G, Zerial M, Di Fiore PP. The eps8 protein coordi-nates egf receptor signalling through rac and trafficking through rab5. Nature, 2000, 408(6810): 374-377.
[6] Miaczynska M, Christoforidis S, Giner A, Shevchenko A, Uttenweiler-Joseph S, Habermann B, Wilm M, Parton RG, Zerial M. Appl proteins link rab5 to nuclear signal trans-duction via an endosomal compartment. Cell, 2004, 116(3): 445-456.
[7] Lanzetti L, Palamidessi A, Areces L, Scita G, Di Fiore PP. Rab5 is a signalling gtpase involved in actin remodelling by receptor tyrosine kinases. Nature, 2004, 429(6989): 309-314.
[8] Eggenschwiler JT, Anderson KV. Dorsal and lateral fates in the mouse neural tube require the cell-autonomous ac-tivity of the open brain gene. Dev Biol, 2000, 227(2): 648-660.
[9] Eggenschwiler JT, Espinoza E, Anderson KV. Rab23 is an essential negative regulator of the mouse sonic hedgehog signalling pathway. Nature, 2001, 412(6843): 194-198.
[10] Guo A, Wang T, Ng EL, Aulia S, Chong KH, Teng FY, Wang Y, Tang BL. Open brain gene product rab23. Ex-pression pattern in the adult mouse brain and functional characterization. J Neurosci Res, 2006, 83(6): 1118-1127.
[11] Eggenschwiler JT, Bulgakov OV, Qin J, Li T, Anderson KV. Mouse rab23 regulates hedgehog signaling from smoothened to gli proteins. Dev Biol, 2006, 290(1): 1-12.
[12] Holland PW. Wholemount in situ hybridization to amphi-oxus embryos. Methods Mol Biol, 1999, 97: 641-644.
[13] Gunther T, Struwe M, Aguzzi A, Schughart K. Open brain, a new mouse mutant with severe neural tube defects, shows altered gene expression patterns in the developing spinal cord. Development, 1994, 120(11): 3119-3130.
[14] Olkkonen VM, Peterson JR, Dupree P, Lutcke A, Zerial M, Simons K. Isolation of a mouse cdna encoding rab23, a small novel gtpase expressed predominantly in the brain. Gene, 1994, 138(1-2): 207-211.
[15] Jenkins D, Seelow D, Jehee FS, Perlyn CA, Alonso LG, Bueno DF, Donnai D, Josifova D, Mathijssen IM, Morton JE, Orstavik KH, Sweeney E, Wall SA, Marsh JL, Nurn-berg P, Passos-Bueno MR, Wilkie AO. Rab23 mutations in carpenter syndrome imply an unexpected role for hedgehog signaling in cranial-suture development and obesity. Am J Hum Genet, 2007, 80(6): 1162-1170.
[16] Evans TM, Ferguson C, Wainwright BJ, Parton RG, Wicking C. Rab23, a negative regulator of hedgehog sig-naling, localizes to the plasma membrane and the endo-cytic pathway. Traffic, 2003, 4(12): 869-884.
[17] Li N, Volff JN, Wizenmann A. Rab23 gtpase is expressed asymmetrically in hensen’s node and plays a role in the dorsoventral patterning of the chick neural tube. Dev Dyn, 2007, 236(11): 2993-3006.
[18] Ng EL, Tang BL. Rab gtpases and their roles in brain neurons and glia. Brain Res Rev, 2008, 58(1): 236-246.
[19] 黄淑红, 杨凌, 谢国蕊, 张红卫. Rab23在小鼠不同器官中表达的研究. 山东大学学报(理学版), 2008, 43(1): 1-4
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