Molecular Characterization and Expression Pattern of Rheb Gene in Inner Mongolia Cashmere Goat (Capra hircus)

doi:10.1016/S1671-2927(11)60138-7

Journal of Integrative Agriculture ›› 2011, Vol. 10 ›› Issue (9): 1452-1458.DOI: 10.1016/S1671-2927(11)60138-7

Molecular Characterization and Expression Pattern of Rheb Gene in Inner Mongolia Cashmere Goat (Capra hircus)

ZHENG Xu, YANG Jiao-fu, WANG Xiao-jing, LIANG Yan, WU Man-lin, SHI Jie-jun, ZHANG Tao, QIN Yin, LI Shu-yu, HAO Xi-yan, WANG Zhi-gang , LIU Dong-jun

College of Life Science, Inner Mongolia University/Key Laboratory of Mammal Reproductive Biology and Biotechnology, Ministry of Education,

收稿日期:2010-08-02 出版日期:2011-09-01 发布日期:2011-09-09
通讯作者: Correspondence WANG Zhi-gang, Professor, Ph D, Mobile: 13190603899, Fax: +86-471-4995867-8008, E-mail:lswzg@imu.edu.cn
作者简介:ZHENG Xu, MSc, E-mail: dong2186@163.com
基金资助:
This wok was supported by grant from the National Natural Science Foundation of China (30860191), the Major Projects for New Varieties of Genetically Modified Organisms, China (2008ZX08008-002) and the Training Fund for the Basic Sciences of China (J0730648).

Molecular Characterization and Expression Pattern of Rheb Gene in Inner Mongolia Cashmere Goat (Capra hircus)

ZHENG Xu, YANG Jiao-fu, WANG Xiao-jing, LIANG Yan, WU Man-lin, SHI Jie-jun, ZHANG Tao, QIN Yin, LI Shu-yu, HAO Xi-yan, WANG Zhi-gang , LIU Dong-jun

College of Life Science, Inner Mongolia University/Key Laboratory of Mammal Reproductive Biology and Biotechnology, Ministry of Education,

Received:2010-08-02 Online:2011-09-01 Published:2011-09-09
Contact: Correspondence WANG Zhi-gang, Professor, Ph D, Mobile: 13190603899, Fax: +86-471-4995867-8008, E-mail:lswzg@imu.edu.cn
About author:ZHENG Xu, MSc, E-mail: dong2186@163.com
Supported by:
This wok was supported by grant from the National Natural Science Foundation of China (30860191), the Major Projects for New Varieties of Genetically Modified Organisms, China (2008ZX08008-002) and the Training Fund for the Basic Sciences of China (J0730648).

摘要/Abstract

摘要： As one member of the Ras super family, Rheb is an upstream regulator of mTOR signaling pathway, which regulates the process of cell-growth, proliferation and differentiation. In order to study the relationship between Rheb and mTOR in Inner Mongolian Cashmere goat (Capra hircus) cells, Ras homolog enriched in brain (Rheb) gene cDNA was amplified by RT-PCR. It is 555 bp in length and includes the complete ORF encoding 184 amino acids (GenBank accession no. HM569224). The full cDNA nucleotide sequence has a 99% identity with that of sheep, 98% with cattle and 93% with human while their amino acids sequence shares identity with 98, 97 and 97% of them, correspondingly. The bioinformatics analysis showed that Rheb has a Ras family domain, two casein kinase II phosphorylation sites, two ATP/GTP-binding sites motif A (P-loop), a prenyl group binding site (CAAX box). Tissue-specific expression analysis performed by semiquantitative RT-PCR. The Rheb gene was expressed in all the tested tissues and the highest level of mRNA accumulation was detected in brain, suggesting that Rheb played an important role in goat cells.

关键词:

Inner Mongolia Cashmere goat, Rheb gene, expression pattern

Abstract: As one member of the Ras super family, Rheb is an upstream regulator of mTOR signaling pathway, which regulates the process of cell-growth, proliferation and differentiation. In order to study the relationship between Rheb and mTOR in Inner Mongolian Cashmere goat (Capra hircus) cells, Ras homolog enriched in brain (Rheb) gene cDNA was amplified by RT-PCR. It is 555 bp in length and includes the complete ORF encoding 184 amino acids (GenBank accession no. HM569224). The full cDNA nucleotide sequence has a 99% identity with that of sheep, 98% with cattle and 93% with human while their amino acids sequence shares identity with 98, 97 and 97% of them, correspondingly. The bioinformatics analysis showed that Rheb has a Ras family domain, two casein kinase II phosphorylation sites, two ATP/GTP-binding sites motif A (P-loop), a prenyl group binding site (CAAX box). Tissue-specific expression analysis performed by semiquantitative RT-PCR. The Rheb gene was expressed in all the tested tissues and the highest level of mRNA accumulation was detected in brain, suggesting that Rheb played an important role in goat cells.

Key words:

Inner Mongolia Cashmere goat, Rheb gene, expression pattern

ZHENG Xu, YANG Jiao-fu, WANG Xiao-jing, LIANG Yan, WU Man-lin, SHI Jie-jun, ZHANG Tao, QIN Yin, LI Shu-yu, HAO Xi-yan, WANG Zhi-gang , LIU Dong-jun . Molecular Characterization and Expression Pattern of Rheb Gene in Inner Mongolia Cashmere Goat (Capra hircus) [J]. Journal of Integrative Agriculture, 2011, 10(9): 1452-1458.

参考文献

[1]Aspuria PJ, Tamanoi F. 2004. The Rheb family of GTP-binding proteins. Cell Singnal, 16, 1105-1112.

[2]Bai X, Ma D, Liu A, Shen X, Wang Q J, Liu Y, Jiang Y. 2007. Rheb activaties mTOR by antagonizing its endogenous inhibitor FKBP38. Science Magazine, 318, 977-980.

[3]Bourne H R, Sanders D A, McCormick F. 1991. The GTPase superfamily: conserved structure and molecular mechanism. Nature, 349, 117-127.

[4]Castro A F, Rebhun J F, Clark G J, Quilliam L A. 2003. Rheb binds tuberous sclerosis complex 2 (TSC2) and promotes S6 kinase activation in a rapamycin- and farnesylation-dependent manner. The Journal of Biological Chemistry, 278, 32493- 32496. Clark G J, Kinch M S, Graham K R, Sebti S M, Hamilton A D, Der C J. 1997. The Ras-related protein Rheb is farnesylated and antagonizes Ras signaling and transformation. The Journal of Biological Chemistry, 272, 10608-10615.

[5]Eom M, Han A, Yi S Y, Shin J J, Cui Y, Park K H. 2008. RHEB expression in fibroadenomas of the breast. Pathology International, 58, 226-232.

[6]Fischer G, Tradler T, Zarnt T. 1998. The mode of action of peptidyl prolyl cis/trans isomerases in vivo: bingding vs. catalysis. FEBS Letters, 426, 17-20.

[7]Hancock J F, Magee A I, Marshall C J. 1989. All ras proteins are polyisoprenylated but only some are palmitoylated. Cell, 57, 1167-1177.

[8]Harrar Y, Bellini C, Faure J D. 2001. FKBPs: at the crossroads of folding and transduction. Trends in Plant Science, 6, 426- 431. Hay H, Sonenberg N. 2004. Upstream and downstream of mTOR. Gene & Development, 18, 1926-1945.

[9]Inoki K, Li Y, Xu T, Guan K L. 2003. Rheb GTPase is a direct target of TSC2 GAP activity and regulates mTOR signaling. Genes & Development, 17, 1829-1834.

[10]Katharina U, Matthias W, Reinhard W, Gunter F, Alfred W, Lgnacio R. 2009. Reassessment of the role of FKBP38 in the Rheb/Mtorc1 pathway. FEBS Letters, 583, 965-970.

[11]Kinsella B T, Erdman R A, Maltese W A. 1991. Posttranslational modification of Ha-ras p21 by farnysyl venrsus geranylgenranyl isoprenoids is determined by the COOHterminal amino acid. Proceedings of the National Academy of Sciences of the USA, 88, 8934-8938.

[12]Knox J P. 1995. The extracellular matrix in higher plants. 4. Developmentally regulated proteoglycans and glycoproteins of the plant cell surface. The FASEB Journal, 9, 1004-1012.

[13]Lau L F, Nathans D. 1987. Expression of a set of growth-related immediate early genes in BALB/c 3T3 cells: Coordinate regulation with c-fos or c-myc. Proceedings of the National Academy of Sciences of the USA, 84, 1182-1186.

[14]Long X, Lin Y, Ortiz-Vega S, Yonezawa K, Avruch J. 2005. Rheb binds and regulates mTOR kinase. Current Biology, 15, 702- 713. Long X, Lin Y, Ortiz-Vega S, Busch S, Avruch J. 2007. The Rheb switch 2 segment is critical for signaling to target of rapamycin complex 1. The Journal of Biological Chemistry, 25, 18542- 18551. Ma D, Bai X, Guo S, Jiang Y. 2008. The switch I region of Rheb is critical for its interaction with FKBP38. The Journal of Biological Chemistry, 283, 25963-25970.

[15]Maestre-Martínez M, Edlich F, Jarczowski F, Weiwad M, Fischer G, Lücke C. 2006. Solution structure of the FK506- binding domain of human FKBP38. Journal of Biomolecular NMR, 34, 197-202.

[16]Nielsen J V, Mitchelmore C, Pedersen K M, Kjaerulff K M, Finsen B, Jensen N A. 2004. Fkbp8: novel isoforms, genomic organization, and characterization of a forebrain promoter in transgenic mice. Genomics, 83, 181-192.

[17]Pedersen K M, Finsen B, Celis J E, Jensen N A. 1999. muFKBP38: a novel murine immunophilin homolog differentially expressed in Schwannoma cells and central nervous system neurons in vivo. Electrophoresis, 20, 249- 255. Saffen D W, Cole A J, Worley P F, Christy B A, Ryder K, Baraban J M. 1988. Convulsant-induced increase in transcription factor messenger RNAs in rat brain. Proceedings of the National Academy of Sciences of the USA, 85, 7795- 7799. Sarbassov D D, Ali S M, Sabatini D M. 2005. Growing role for the mTOR pathway. Current Opinion in Cell Biology, 17, 596-607.

[18]Sato T, Umetsu A, Tamanoi F. 2008. Characterization of the Rheb-mTOR signaling pathway in mammalia cells: constitietive active mutants of Rheb and mTOR. Methods Enzymol, 438, 307-320.

[19]Shirane M, Nakayama K I. 2003. Inherent calcineurin inhibitor FKBP38 targets Bcl-2 to mitochondria and inhibits apoptosis. Nature Cell Biology, 5, 28-37.

[20]Tee A R, Manning B D, Roux P P, Cantley L C, Blenis J. 2003. Tuberous sclerosis complex gene products, tuberin and hamartin, control mTOR signaling by acting as a GTPaseactivating protein complex toward Rheb. Current Biology, 13, 1259-1268.

[21]Tee A R, Blenis J, Proud C G. 2005. Analysis of mTOR signaling by the small G-proteins, Rheb and RhebL1. FEBS Letters, 29, 4763-4768.

[22]Urano J, Tabancay A P, Yang W, Tamanoi F. 2000. The Saccharomyces cerevisiae Rheb G-protein is involved in regulating canavanine resistance and arginine uptake. The Journal of Biological Chemistry, 275, 11198-11206.

[23]Wang H Q, Nakaya Y, Du Z, Yamane T, Shirane M, Kudo T, Takeda M, Takebayashi K, Noda Y, Nakayama K I, et al. 2005. Interaction of presenilins with FKBP38 promotes apoptosis by reducing mitochondrial Bcl-2. Human Molecular Genetics, 14, 1889-1902.

[24]Yamagata K, Sanders L K, Kaufmann W E, Yee W, Barnes C A, Nathans D, Worley P F. 1994. Rheb, a growth factor-and synaptic-regulated gene, encodes a novel Ras-related protein. The Journal of Biological Chemistry, 269, 16333-16339.

[25]Zhang Y, Gao X, Saucedo L J, Ru B, Edgar B A, Pan D. 2003. Rheb is a direct target of the tuberous sclerosis tumour suppressor proteins. Nature Cell Biology, 5, 578-581.

Molecular Characterization and Expression Pattern of Rheb Gene in Inner Mongolia Cashmere Goat (Capra hircus)

Molecular Characterization and Expression Pattern of Rheb Gene in Inner Mongolia Cashmere Goat (Capra hircus)

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