Abstract
Smad family proteins are identified as intracellular signal mediators of the TGF-β superfamily. In this study, we identified two novel members of the Smad family, termed as AmphiSmad1/5/8 and AmphiSmad4, from Chinese amphioxus. Both AmphiSmad1/5/8 and AmphiSmad4 showed a typical domain structure of Smad proteins consisting of conserved MH1 and MH2 domains. Phylogenetic analysis placed AmphiSmad1/5/8 in the Smad1, 5 and 8 subgroup of the R-Smad subfamily, and AmphiSmad4 in the Co-Smad subfamily. The spatial and temporal gene expression patterns of AmphiSmad1/5/8 and AmphiSmad4 showed that they may be involved in the embryonic development of notochord, myotome and alimentary canal, and may help to establish the specification of dorsal-ventral axis of amphioxus. Moreover, AmphiSmad1/5/8 and AmphiSmad4 showed extensive distribution in all adult tissues examined, suggesting that these two genes may play important roles in the morphogenesis of a variety of tissues especially notochord and gonad.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Massagué J, Blain S W, Lo R S. TGF-β signaling in growth control, cancer, and heritable disorders. Cell, 2000, 103: 295–309, 11057902, 10.1016/S0092-8674(00)00121-5
Massagué J, Seoane J, Wotton D. Smad transcription factors. Genes Dev, 2005, 19: 2783–2810, 16322555, 10.1101/gad.1350705
Raftery L A, Twombly V, Wharton K, et al. Genetic screens to identify elements of the decapentaplegic signaling pathway in Drosophila. Genetics, 1995, 139: 241–254, 7705627, 1:CAS:528:DyaK28Xht1Wqu7Y%3D
Sekelsky J J, Newfeld S J, Raftery L A, et al. Genetic characterization and cloning of mothers against dpp, a gene required for decapentaplegic function in Drosophila melanogaster. Genetics, 1995, 139: 1347–1358, 7768443, 1:CAS:528:DyaK28XhsFSrurw%3D
Shi Y, Massagué J. Mechanisms of TGF-β signaling from cell membrane to the nucleus. Cell, 2003, 113: 685–700, 12809600, 10.1016/S0092-8674(03)00432-X, 1:CAS:528:DC%2BD3sXkvVekt7o%3D
Inman G J, Nicolas F J, Hill C S. Nucleocytoplasmic shuttling of Smads 2, 3 and 4 permits sensing of TGF-β receptor activity. Mol Cell, 2002, 10: 283–294, 12191474, 10.1016/S1097-2765(02)00585-3, 1:CAS:528:DC%2BD38Xmslyrtbg%3D
Xu L, Massagué J. Nucleocytoplasmic shuttling of signal transducers. Nat Rev Mol Cell Biol, 2004, 5: 209–219, 14991001, 10.1038/nrm1331, 1:CAS:528:DC%2BD2cXhs1Clt7k%3D
Miyazono K, Ten D P, Heldin C H. TGF-beta signaling by Smad proteins. Adv Immunol, 2000, 75: 115–157, 10879283, 10.1016/S0065-2776(00)75003-6, 1:CAS:528:DC%2BD3cXltFSru7c%3D
Weinstein M, Yang X, Deng C. Function of mammalian Smad genes as revealed by targeted gene disruption in mice. Cytokine Growth Factor Rev, 2000, 11: 49–58, 10708952, 10.1016/S1359-6101(99)00028-3, 1:CAS:528:DC%2BD3cXitlGjsrw%3D
Arnold S J, Maretto S, Islam A, et al. Dose-dependent Smad1, Smad5 and Smad8 signaling in the early mouse embryo. Dev Biol, 2006, 296: 104–118, 16765933, 10.1016/j.ydbio.2006.04.442, 1:CAS:528:DC%2BD28XntV2mur0%3D
Lau M T, Ge W. Cloning of Smad2, Smad3, Smad4, and Smad7 from the goldfish pituitary and evidence for their involvement in activin regulation of goldfish FSHβ promoter activity. Gen Comp Endocrinol, 2005, 141: 22–38, 15707600, 10.1016/j.ygcen.2004.10.019, 1:CAS:528:DC%2BD2MXhtFGhtb8%3D
Holland L Z, Laudet V, Schubert M. The chordate amphioxus: An emerging model organism for developmental biology. Cell Mol Life Sci, 2004, 61: 2290–308, 15378201, 10.1007/s00018-004-4075-2, 1:CAS:528:DC%2BD2cXhtVWlu7nM
Holland P W. Whole mount in situ hybridization to amphioxus embryos. Methods Mol Bio, 1999, 97: 641–644, 1:CAS:528:DyaK1MXltVKmsb4%3D
Graff J M, Bansal A, Melton D A. Xenopus Mad proteins transducer distinct subsets of signals for the TGF-β superfamily. Cell, 1996, 85: 479–487, 8653784, 10.1016/S0092-8674(00)81249-0, 1:CAS:528:DyaK28XjtFyitrY%3D
Thomsen G H. Xenopus mothers against decapentaplegic is an embryonic ventralizing agent that acts downstream of the BMP-2/4 receptor. Development, 1996, 122: 2359–2366, 8756281, 1:CAS:528:DyaK28XltFWnt78%3D
Suzuki A, Chang C B, Yingling J M, et al. Hemmati-Brivanlou, Smad5 induces ventral fates in Xenopus embryo. Dev Biol, 1997, 184: 402–405, 9133445, 10.1006/dbio.1997.8548, 1:CAS:528:DyaK2sXjtVaht7Y%3D
Baker J C, Harland R M. A novel mesoderm inducer, Madr2, functions in the activin signal transduction pathway. Genes Dev, 1996, 10: 1880–1889, 8756346, 10.1101/gad.10.15.1880, 1:CAS:528:DyaK28XkvFGlt7Y%3D
Muller F, Blader P, Rastegar S, et al. Characterization of zebrafish smad1, smad2 and smad5: The amino-terminus of Smad1 and Smad5 for specific function in the embryo. Mech Dev, 1999, 88: 73–88, 10525190, 10.1016/S0925-4773(99)00173-2, 1:CAS:528:DyaK1MXmsFaru7s%3D
Panopoulou G, Clark M D, Holland L Z, et al. AmphiBMP2/4, an amphioxus bone morphogenetic protein closely related to Drosophila decapentaplegic and veterbrate BMP2 and BMP4: Insights into evolution of dorsoventral axis specification. Dev Dyn, 1998, 213: 130–139, 9733108, 10.1002/(SICI)1097-0177(199809)213:1<130::AID-AJA13>3.0.CO;2-6, 1:CAS:528:DyaK1cXlvF2iu7Y%3D
Yu J K, Satou Y, Holland N D, et al. Axial patterning in cephalochordates and the evolution of the organizer. Nature, 2007, 445: 613–617, 17237766, 10.1038/nature05472, 1:CAS:528:DC%2BD2sXhtlOrtLo%3D
Yasui K, Saiga H, Wang Y, et al. Early expressed genes showing a dichotomous developing pattern in the lancelet embryo. Dev Growth Differ, 2001, 43: 185–194, 11284968, 10.1046/j.1440-169X.2001.00566.x, 1:CAS:528:DC%2BD3MXktValsLw%3D
Yang X, Li C, Xu X, et al. The tumor suppressor Smad4/DPC4 is essential for epiblast proliferation and mesoderm induction in mice. Proc Natl Acad Sci USA, 1998, 95: 3667–3672, 9520423, 10.1073/pnas.95.7.3667, 1:CAS:528:DyaK1cXitlKjtr8%3D
Dick A, Mayr T, Bauer H, et al. Cloning and characterization of zebrafish smad2, smad3 and smad4. Gene, 2000, 246: 69–80, 10767528, 10.1016/S0378-1119(00)00056-1, 1:CAS:528:DC%2BD3cXitleku7w%3D
Flanders K C, Kim E S, Roberts A B. Immunohistochemical expression of Smads 1–6 in the 15 day gestatin mouse embryo: Signaling by BMPs and TGF-βs. Dev Dyn, 2001, 200: 141–154, 10.1002/1097-0177(2000)9999:9999<::AID-DVDY1096>3.0.CO;2-4
Luukko K, Ylikorkala A, Makela T P. Developmentally regulated expression of Smad3, Smad4, Smad6, and Smad7 involved in TGF-beta signaling. Mech Dev, 2001, 101: 209–212, 11231077, 10.1016/S0925-4773(00)00556-6, 1:CAS:528:DC%2BD3MXhsFShtbk%3D
Howell M, Itoh F, Pierreus C E, et al. Xenpous Smad4β is the Co-Smad component of developmentally regulated transcription factor complexes responsible for induction of early mesodermal genes. Dev Biol, 1999, 214: 354–369, 10525340, 10.1006/dbio.1999.9430, 1:CAS:528:DyaK1MXms1CrtLs%3D
Masuyama N, Hanafusa H, Kusakabe M, et al. Identification of two Smad4 proteins in Xenopus. J Biol Chem, 1999, 274: 12163–12170, 10207044, 10.1074/jbc.274.17.12163, 1:CAS:528:DyaK1MXislGltLo%3D
Wang Z Y, Futami K, Nishihara A, et al. Four types of Smad4 found in the common carp, Cyprinus carpio. J Exp Zoolog B Mol Dev Evol, 2005, 304: 250–258, 10.1002/jez.b.21041
Huang S X, Flanders K C, Roberts A B. Characterization of the mouse Smad1 gene and its expression pattern in adult mouse tissues. Gene, 2000, 258: 43–53, 11111041, 10.1016/S0378-1119(00)00396-6, 1:CAS:528:DC%2BD3cXosFejuro%3D
Howell M, Mohun T J, Hill C S. Xenopus Smad3 is specifically expressed in the chordoneural henge, notochord and in the endocardium of the developing heart. Mech Dev, 2001, 104: 147–150, 11404094, 10.1016/S0925-4773(01)00365-3, 1:CAS:528:DC%2BD3MXkt1yjuro%3D
Oxbrugh L, Robertson E J. Dynamic regulation of Smad expression during mesenchyme to epithelium transition in the metanephric kidney. Mec Dyn, 2002, 12: 207–211
Kaivo-oja N, Jeffery L A, Ritvos O, et al. Smad signalling in the ovary. Reprod Biol Endocrinol, 2006, 12: 14–21
Vargesson N, Laufer E. Negative Smad expression and regulation in the developing chick limb. PLoS ONE, 2009, 4: 1–8, 10.1371/journal.pone.0005173
Author information
Authors and Affiliations
Corresponding authors
Additional information
Contributed equally to this work
Rights and permissions
Open Access This is an open access article distributed under the terms of the Creative Commons Attribution Noncommercial License ( https://creativecommons.org/licenses/by-nc/2.0 ), which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.
About this article
Cite this article
Yu, X., Li, J., Liu, H. et al. Identification and expression of amphioxus AmphiSmad1/5/8 and AmphiSmad4. Sci. China Life Sci. 54, 220–226 (2011). https://doi.org/10.1007/s11427-011-4136-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11427-011-4136-3