槟榔江水牛ACVR1基因分离鉴定及表达谱分析
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摘要
【目的】ACVR1作为一些TGF-β超家族成员信号通路的I型受体之一,在器官形成和细胞分化及卵泡形成等调控事件中扮演重要角色。迄今,有关ACVR1基因的研究主要集中在人上,有关水牛ACVR1基因的研究尚未见报道。【方法】采用RT-PCR测序法对槟榔江水牛ACVR1基因的编码区序列进行了分离鉴定,进一步对之进行了功能生物信息学和表达谱分析。【结果】水牛ACVR1基因编码区全长1 530 bp,编码1个由509个氨基酸残基构成的多肽。水牛ACVR1为弱亲水性蛋白,含有1个信号肽和跨膜区,定位在质膜上;该蛋白含有Activin_recp、TGF_beta_GS和STKc_ACVR1_ALK1三个保守结构域,其二级结构、三维结构与人的ACVR1极为相似。水牛的ACVR1与普通牛、野牦牛、人和猪等8个物种的ACVR1氨基酸序列间一致性99%。在所检测的水牛9个组织中,ACVR1基因在肝脏、脾脏、肾、瘤胃、卵巢、子宫和睾丸中均表达,其中在卵巢和子宫中表达水平最高。【结论】水牛的ACVR1与其他物种的ACVR1具有相近的氨基酸组成和结构特征,其作为BMP、AMH信号通路的I型共享受体,可能参与了水牛的细胞分化及卵泡形成等调控事件。
[Purpose] ACVR1, as one of the type I receptors of some members of TGF-β superfamily signaling pathways, plays an important role in the regulatory events such as organ formation,cell differentiation and follicle formation. Up to now, the studies on ACVR1 gene mainly focuses on human, and the research on this gene in water buffalo has not been reported yet.[Methods]In this study, the coding region of ACVR1 gene of Binglangjiang buffalo was isolated and identified by RTPCR method, and the gene was then subjected to analysis of functional bioinformatics and gene expression profiling.[Results]The full coding region of buffalo ACVR1 gene is 1 530 bp in length,which encodes a polypeptide consisting of 509 amino acid residues. Buffalo ACVR1 is a weakly hydrophilic protein containing a signal peptide and transmembrane region located on the plasma membrane. Structurally, buffalo ACVR1 contains three conserved domains of Activin_recp, TGF_beta_GS and STKc_ACVR1_ALK1. The secondary structure, three-dimensional structure of this protein are very similar to human ACVR1. The consistency of ACVR1 amino acid sequences between buffalo and 8 other common species such as cattle, wild yak, human and pig was 99%. Among the 9 buffalo tissues assayed, the ACVR1 gene was expressed in the liver, spleen, kidney, rumen, ovary, uterus and testes with that the highest expression in the ovary and uterus.[Conclusion]The buffalo ACVR1 has similar amino acid composition and structural characteristics with that of other species. As one of type I receptor shared by BMP and AMH signaling pathway, ACVR1 may be involved in the regulation of buffalo cell differentiation and follicular formation.
[Purpose] ACVR1, as one of the type I receptors of some members of TGF-β superfamily signaling pathways, plays an important role in the regulatory events such as organ formation,cell differentiation and follicle formation. Up to now, the studies on ACVR1 gene mainly focuses on human, and the research on this gene in water buffalo has not been reported yet.[Methods]In this study, the coding region of ACVR1 gene of Binglangjiang buffalo was isolated and identified by RTPCR method, and the gene was then subjected to analysis of functional bioinformatics and gene expression profiling.[Results]The full coding region of buffalo ACVR1 gene is 1 530 bp in length,which encodes a polypeptide consisting of 509 amino acid residues. Buffalo ACVR1 is a weakly hydrophilic protein containing a signal peptide and transmembrane region located on the plasma membrane. Structurally, buffalo ACVR1 contains three conserved domains of Activin_recp, TGF_beta_GS and STKc_ACVR1_ALK1. The secondary structure, three-dimensional structure of this protein are very similar to human ACVR1. The consistency of ACVR1 amino acid sequences between buffalo and 8 other common species such as cattle, wild yak, human and pig was 99%. Among the 9 buffalo tissues assayed, the ACVR1 gene was expressed in the liver, spleen, kidney, rumen, ovary, uterus and testes with that the highest expression in the ovary and uterus.[Conclusion]The buffalo ACVR1 has similar amino acid composition and structural characteristics with that of other species. As one of type I receptor shared by BMP and AMH signaling pathway, ACVR1 may be involved in the regulation of buffalo cell differentiation and follicular formation.
引文
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[15]哈福,李大林,袁跃云,等.牛科物种FSHβ基因外显子1和外显子2遗传特征分析[J].云南农业大学学报(自然科学),2013,28(1):61.DOI:10.3969/j.issn.1004-390X(n).2013.01.011.
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[17]FENG X H,DERYNCK R.A kinase subdomain of transforming growth factor-beta(TGF-beta)type I receptor determines the TGF-beta intracellular signaling specificity[J].The EMBO Journal,1997,16(13):3912.DOI:10.1093/emboj/16.13.3912.
[18]KAPLAN F S,PIGNOLO R J,SHORE E M.The FOPmetamorphogene encodes a novel type I receptor that dysregulates BMP signaling[J].Cytokine&Growth Factor Reviews,2009,20(5/6):399.DOI:10.1016/j.cyto gfr.2009.10.006.
[19]KAPLAN F S,SHEN Q,LOUNEV V,et al.Skeletal metamorphosis in fibrodysplasia ossificans progressiva(FOP)[J].Journal of Bone and Mineral Metabolism,2008,26(6):521.DOI:10.1007/s00774-008-0879-8.
[20]VAN DINTHER M,VISSER N,DE GORTER D J,et al.ALK2 R206H mutation linked to fibrodysplasia ossificans progressiva confers constitutive activity to the BMPtype I receptor and sensitizes mesenchymal cells to BMP-induced osteoblast differentiation and bone formation[J].Journal of Bone and Mineral Research,2010,25(6):1208.DOI:10.1359/jbmr.091110.
[21]SHIMASAKI S,MOORE R K,OTSUKA F,et al.The bone morphogenetic protein system in mammalian reproduction[J].Endocrine Reviews,2004,25(1):72.DOI:10.1210/er.2003-0007.
[22]KEVENAAR M E,THEMMEN A P,VAN KERKWIJKA J,et al.Variants in the ACVR1 gene are associated with AMH levels in women with polycystic ovary syndrome[J].Human Reproduction,2009,24(1):241.DOI:10.1093/humrep/den353.
[2]CLáUDIA A S.ACVR1-activin A receptor,type I[J].Atlas of Genetics and Cytogenetics in Oncology and Hae matology,2013,17(10):670.DOI:10.4267/2042/51532.
[3]SHORE E M,XU M Q,FELDMAN G J,et al.A recurrent mutation in the BMP type I receptor ACVR1 causes inherited and sporadic fibrodysplasia ossificans progressiva[J].Nature Genetics,2006,38(5):525.DOI:10.1038/ng1783.
[4]DANDARA C,SCOTT C,URBAN M,et al.Confirmation of the recurrent ACVR1 617G>A mutation in South Africans with fibrodysplasia ossificans progressiva[J].South African Medical Journal,2012,102(7):631.DOI:10.7196/SAMJ.5519.
[5]PIGNOLO R J,SHORE E M,KAPLAN F S.Fibrodysplasia ossificans progressiva:diagnosis,management,and therapeutic horizons[J].Pediatric Endocrinology Reviews:PER,2013,10(S2):437.
[6]ERESEN YAZICIO?LU C,KARATOSUN V,KIZ-ILDA?S,et al.ACVR1 gene mutations in four Turkish patients diagnosed as fibrodysplasia ossificans progressiva[J].Gene,2013,515(2):444.DOI:10.1016/j.gene.2012.12.005.
[7]ERICKSON G F,SHIMASAKI S.The spatiotemporal expression pattern of the bone morphogenetic protein family in rat ovary cell types during the estrous cycle[J].Reproductive Biology and Endocrinology,2003,1(1):9.
[8]NILSSON E E,SKINNER M K.Bone morphogenetic protein-4 acts as an ovarian follicle survival factor and promotes primordial follicle development[J].Biology of Reproduction,2003,69(4):1265.DOI:10.1095/biolreprod.103.018671.
[9]VISSER J A.AMH signaling:from receptor to target gene[J].Molecular and Cellular Endocrinology,2003,211(1/2):65.
[10]DI CLEMENTE N,JOSSO N,GOUéDARD L,et al.Components of the anti-Müllerian hormone signaling pathway in gonads[J].Molecular and Cellular Endocrinology,2003,211(1/2):9.
[11]高先栋.猪腹壁(腹膜)骨化症初报[J].畜牧与饲料科学,2010,31(3):177.DOI:10.3969/j.issn.1672-5190.2010.03.088.
[12]ZHAO X,MO D L,LI A N,et al.Comparative analyses by sequencing of transcriptomes during skeletal muscle development between pig breeds differing in muscle growth rate and fatness[J].PLoS One,2011,6(5):e19774.DOI:10.1371/journal.pone.0019774.
[13]王文浩,王金玉,张跟喜,等.ACVR1基因对京海黄鸡生长和繁殖性状的遗传效应研究[J].中国畜牧兽医,2012,39(7):70.DOI:10.3969/j.issn.1671-7236.2012.07.016.
[14]MICHELIZZI V N,DODSON M V,PAN Z A,et al.Water buffalo genome science comes of age[J].International Journal of Biological Sciences,2010,6(4):333.
[15]哈福,李大林,袁跃云,等.牛科物种FSHβ基因外显子1和外显子2遗传特征分析[J].云南农业大学学报(自然科学),2013,28(1):61.DOI:10.3969/j.issn.1004-390X(n).2013.01.011.
[16]KINGSLEY D M.The TGF-beta superfamily:new members,new receptors,and new genetic tests of function in different organisms[J].Genes&Development,1994,8(2):133.
[17]FENG X H,DERYNCK R.A kinase subdomain of transforming growth factor-beta(TGF-beta)type I receptor determines the TGF-beta intracellular signaling specificity[J].The EMBO Journal,1997,16(13):3912.DOI:10.1093/emboj/16.13.3912.
[18]KAPLAN F S,PIGNOLO R J,SHORE E M.The FOPmetamorphogene encodes a novel type I receptor that dysregulates BMP signaling[J].Cytokine&Growth Factor Reviews,2009,20(5/6):399.DOI:10.1016/j.cyto gfr.2009.10.006.
[19]KAPLAN F S,SHEN Q,LOUNEV V,et al.Skeletal metamorphosis in fibrodysplasia ossificans progressiva(FOP)[J].Journal of Bone and Mineral Metabolism,2008,26(6):521.DOI:10.1007/s00774-008-0879-8.
[20]VAN DINTHER M,VISSER N,DE GORTER D J,et al.ALK2 R206H mutation linked to fibrodysplasia ossificans progressiva confers constitutive activity to the BMPtype I receptor and sensitizes mesenchymal cells to BMP-induced osteoblast differentiation and bone formation[J].Journal of Bone and Mineral Research,2010,25(6):1208.DOI:10.1359/jbmr.091110.
[21]SHIMASAKI S,MOORE R K,OTSUKA F,et al.The bone morphogenetic protein system in mammalian reproduction[J].Endocrine Reviews,2004,25(1):72.DOI:10.1210/er.2003-0007.
[22]KEVENAAR M E,THEMMEN A P,VAN KERKWIJKA J,et al.Variants in the ACVR1 gene are associated with AMH levels in women with polycystic ovary syndrome[J].Human Reproduction,2009,24(1):241.DOI:10.1093/humrep/den353.