The PTPN11 (protein-tyrosine phosphatase, non-receptor type 11) gene encodes SHP2, a cytoplasmic PTP that is essential for vertebrate development. Mutations in PTPN11 are associated with Noonan and LEOPARD syndrome. Human patients with these autosomal dominant disorders display various symptoms, including short stature, craniofacial defects and heart abnormalities. We have used the zebrafish as a model to investigate the role of Shp2 in embryonic development. The zebrafish genome encodes two ptpn11 genes, ptpn11a and ptpn11b. Here, we report that ptpn11a is expressed constitutively and ptpn11b expression is strongly upregulated during development. In addition, the products of both ptpn11 genes, Shp2a and Shp2b, are functional. Target-selected inactivation of ptpn11a and ptpn11b revealed that double homozygous mutants are embryonic lethal at 5–6 days post fertilization (dpf). Ptpn11a-/-ptpn11b-/- embryos showed pleiotropic defects from 4 dpf onwards, including reduced body axis extension and craniofacial defects, which was accompanied by low levels of phosphorylated Erk at 5 dpf. Interestingly, defects in homozygous ptpn11a-/- mutants overlapped with defects in the double mutants albeit they were milder, whereas ptpn11b-/- single mutants did not show detectable developmental defects and were viable and fertile. Ptpn11a-/-ptpn11b-/- mutants were rescued by expression of exogenous ptpn11a and ptpn11b alike, indicating functional redundance of Shp2a and Shp2b. The ptpn11 mutants provide a good basis for further unravelling of the function of Shp2 in vertebrate development.
References
[1]
Dance M, Montagner A, Salles JP, Yart A, Raynal P (2008) The molecular functions of Shp2 in the Ras/Mitogen-activated protein kinase (ERK1/2) pathway. Cell Signal 20: 453–459. doi: 10.1016/j.cellsig.2007.10.002
[2]
Feng GS (1999) Shp-2 tyrosine phosphatase: signaling one cell or many. Exp Cell Res 253: 47–54. doi: 10.1006/excr.1999.4668
[3]
Neel BG, Gu H, Pao L (2003) The ‘Shp’ing news: SH2 domain-containing tyrosine phosphatases in cell signaling. Trends Biochem Sci 28: 284–293. doi: 10.1016/s0968-0004(03)00091-4
[4]
Tartaglia M, Mehler EL, Goldberg R, Zampino G, Brunner HG, et al. (2001) Mutations in PTPN11, encoding the protein tyrosine phosphatase SHP-2, cause Noonan syndrome. Nat Genet 29: 465–468.
[5]
Legius E, Schrander-Stumpel C, Schollen E, Pulles-Heintzberger C, Gewillig M, et al. (2002) PTPN11 mutations in LEOPARD syndrome. J Med Genet 39: 571–574. doi: 10.1136/jmg.39.8.571
[6]
Digilio MC, Conti E, Sarkozy A, Mingarelli R, Dottorini T, et al. (2002) Grouping of multiple-lentigines/LEOPARD and Noonan syndromes on the PTPN11 gene. Am J Hum Genet 71: 389–394. doi: 10.1086/341528
[7]
Martinez-Quintana E, Rodriguez-Gonzalez F (2013) RASopathies: From Noonan to LEOPARD Syndrome. Rev Esp Cardiol.
[8]
Rodriguez-Viciana P, Tetsu O, Tidyman WE, Estep AL, Conger BA, et al. (2006) Germline mutations in genes within the MAPK pathway cause cardio-facio-cutaneous syndrome. Science 311: 1287–1290. doi: 10.1126/science.1124642
[9]
Zenker M (2009) Noonan Syndrome and Related Disorders. A matter of Deregulated Ras Signaling.; Schmid M, editor. Basel, Switzerland: Karger. 24 p.
[10]
Tartaglia M, Zampino G, Gelb BD (2010) Noonan syndrome: clinical aspects and molecular pathogenesis. Mol Syndromol 1: 2–26. doi: 10.1159/000276766
[11]
Hanna N, Montagner A, Lee WH, Miteva M, Vidal M, et al. (2006) Reduced phosphatase activity of SHP-2 in LEOPARD syndrome: consequences for PI3K binding on Gab1. FEBS Lett 580: 2477–2482. doi: 10.1016/j.febslet.2006.03.088
[12]
Kontaridis MI, Swanson KD, David FS, Barford D, Neel BG (2006) PTPN11 (Shp2) mutations in LEOPARD syndrome have dominant negative, not activating, effects. J Biol Chem 281: 6785–6792. doi: 10.1074/jbc.m513068200
[13]
Tartaglia M, Martinelli S, Stella L, Bocchinfuso G, Flex E, et al. (2006) Diversity and functional consequences of germline and somatic PTPN11 mutations in human disease. Am J Hum Genet 78: 279–290. doi: 10.1086/499925
[14]
Yu ZH, Xu J, Walls CD, Chen L, Zhang S, et al. (2013) Structural and mechanistic insights into LEOPARD syndrome-associated SHP2 mutations. J Biol Chem 288: 10472–10482. doi: 10.1074/jbc.m113.450023
[15]
Keilhack H, David FS, McGregor M, Cantley LC, Neel BG (2005) Diverse biochemical properties of Shp2 mutants. Implications for disease phenotypes. J Biol Chem 280: 30984–30993. doi: 10.1074/jbc.m504699200
[16]
Oishi K, Gaengel K, Krishnamoorthy S, Kamiya K, Kim IK, et al. (2006) Transgenic Drosophila models of Noonan syndrome causing PTPN11 gain-of-function mutations. Hum Mol Genet 15: 543–553. doi: 10.1093/hmg/ddi471
[17]
Oishi K, Zhang H, Gault WJ, Wang CJ, Tan CC, et al. (2009) Phosphatase-defective LEOPARD syndrome mutations in PTPN11 gene have gain-of-function effects during Drosophila development. Hum Mol Genet 18: 193–201. doi: 10.1093/hmg/ddn336
[18]
Edouard T, Combier JP, Nedelec A, Bel-Vialar S, Metrich M, et al. (2010) Functional effects of PTPN11 (SHP2) mutations causing LEOPARD syndrome on epidermal growth factor-induced phosphoinositide 3-kinase/AKT/glycogen synthase kinase 3beta signaling. Mol Cell Biol 30: 2498–2507. doi: 10.1128/mcb.00646-09
[19]
De Rocca Serra-Nedelec A, Edouard T, Treguer K, Tajan M, Araki T, et al. (2012) Noonan syndrome-causing SHP2 mutants inhibit insulin-like growth factor 1 release via growth hormone-induced ERK hyperactivation, which contributes to short stature. Proc Natl Acad Sci U S A 109: 4257–4262. doi: 10.1073/pnas.1119803109
[20]
Yang W, Klaman LD, Chen B, Araki T, Harada H, et al. (2006) An Shp2/SFK/Ras/Erk signaling pathway controls trophoblast stem cell survival. Dev Cell 10: 317–327. doi: 10.1016/j.devcel.2006.01.002
[21]
Ke Y, Zhang EE, Hagihara K, Wu D, Pang Y, et al. (2007) Deletion of Shp2 in the brain leads to defective proliferation and differentiation in neural stem cells and early postnatal lethality. Mol Cell Biol 27: 6706–6717. doi: 10.1128/mcb.01225-07
[22]
Kontaridis MI, Yang W, Bence KK, Cullen D, Wang B, et al. (2008) Deletion of Ptpn11 (Shp2) in cardiomyocytes causes dilated cardiomyopathy via effects on the extracellular signal-regulated kinase/mitogen-activated protein kinase and RhoA signaling pathways. Circulation 117: 1423–1435. doi: 10.1161/circulationaha.107.728865
[23]
Bard-Chapeau EA, Yuan J, Droin N, Long S, Zhang EE, et al. (2006) Concerted functions of Gab1 and Shp2 in liver regeneration and hepatoprotection. Mol Cell Biol 26: 4664–4674. doi: 10.1128/mcb.02253-05
[24]
Bard-Chapeau EA, Li S, Ding J, Zhang SS, Zhu HH, et al. (2011) Ptpn11/Shp2 acts as a tumor suppressor in hepatocellular carcinogenesis. Cancer Cell 19: 629–639. doi: 10.1016/j.ccr.2011.03.023
[25]
Tang TL, Freeman RM Jr, O'Reilly AM, Neel BG, Sokol SY (1995) The SH2-containing protein-tyrosine phosphatase SH-PTP2 is required upstream of MAP kinase for early Xenopus development. Cell 80: 473–483. doi: 10.1016/0092-8674(95)90498-0
[26]
Jopling C, van Geemen D, den Hertog J (2007) Shp2 knockdown and Noonan/LEOPARD mutant Shp2-induced gastrulation defects. PLoS Genet 3: e225. doi: 10.1371/journal.pgen.0030225
[27]
Stewart RA, Sanda T, Widlund HR, Zhu S, Swanson KD, et al. (2010) Phosphatase-dependent and -independent functions of Shp2 in neural crest cells underlie LEOPARD syndrome pathogenesis. Dev Cell 18: 750–762. doi: 10.1016/j.devcel.2010.03.009
[28]
Lieschke GJ, Currie PD (2007) Animal models of human disease: zebrafish swim into view. Nat Rev Genet 8: 353–367. doi: 10.1038/nrg2091
[29]
Kimmel CB, Ballard WW, Kimmel SR, Ullmann B, Schilling TF (1995) Stages of embryonic development of the zebrafish. Dev Dyn 203: 253–310. doi: 10.1002/aja.1002030302
[30]
Wienholds E, van Eeden F, Kosters M, Mudde J, Plasterk RH, et al. (2003) Efficient target-selected mutagenesis in zebrafish. Genome Res 13: 2700–2707. doi: 10.1101/gr.1725103
[31]
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25: 402–408. doi: 10.1006/meth.2001.1262
[32]
Thisse C, Thisse B, Schilling TF, Postlethwait JH (1993) Structure of the zebrafish snail1 gene and its expression in wild-type, spadetail and no tail mutant embryos. Development 119: 1203–1215.
[33]
van Eekelen M, Overvoorde J, van Rooijen C, den Hertog J (2010) Identification and expression of the family of classical protein-tyrosine phosphatases in zebrafish. PLoS One 5: e12573. doi: 10.1371/journal.pone.0012573
[34]
Vandepoele K, De Vos W, Taylor JS, Meyer A, Van de Peer Y (2004) Major events in the genome evolution of vertebrates: paranome age and size differ considerably between ray-finned fishes and land vertebrates. Proc Natl Acad Sci U S A 101: 1638–1643. doi: 10.1073/pnas.0307968100
[35]
Jaillon O, Aury JM, Brunet F, Petit JL, Stange-Thomann N, et al. (2004) Genome duplication in the teleost fish Tetraodon nigroviridis reveals the early vertebrate proto-karyotype. Nature 431: 946–957. doi: 10.1038/nature03025
[36]
Nadler MJ, Chen B, Anderson JS, Wortis HH, Neel BG (1997) Protein-tyrosine phosphatase SHP-1 is dispensable for FcgammaRIIB-mediated inhibition of B cell antigen receptor activation. J Biol Chem 272: 20038–20043. doi: 10.1074/jbc.272.32.20038
[37]
Saxton TM, Henkemeyer M, Gasca S, Shen R, Rossi DJ, et al. (1997) Abnormal mesoderm patterning in mouse embryos mutant for the SH2 tyrosine phosphatase Shp-2. EMBO J 16: 2352–2364. doi: 10.1093/emboj/16.9.2352
[38]
Noonan JA, Raaijmakers R, Hall BD (2003) Adult height in Noonan syndrome. Am J Med Genet A 123A: 68–71. doi: 10.1002/ajmg.a.20502
[39]
Gorlin RJ, Anderson RC, Blaw M (1969) Multiple lentigenes syndrome. Am J Dis Child 117: 652–662.
[40]
Araki T, Mohi MG, Ismat FA, Bronson RT, Williams IR, et al. (2004) Mouse model of Noonan syndrome reveals cell type- and gene dosage-dependent effects of Ptpn11 mutation. Nat Med 10: 849–857. doi: 10.1038/nm1084
[41]
Marin TM, Keith K, Davies B, Conner DA, Guha P, et al. (2011) Rapamycin reverses hypertrophic cardiomyopathy in a mouse model of LEOPARD syndrome-associated PTPN11 mutation. J Clin Invest 121: 1026–1043. doi: 10.1172/jci44972
[42]
Noonan JA (1968) Hypertelorism with Turner phenotype. A new syndrome with associated congenital heart disease. Am J Dis Child 116: 373–380. doi: 10.1001/archpedi.1968.02100020377005
[43]
Tartaglia M, Kalidas K, Shaw A, Song X, Musat DL, et al. (2002) PTPN11 mutations in Noonan syndrome: molecular spectrum, genotype-phenotype correlation, and phenotypic heterogeneity. Am J Hum Genet 70: 1555–1563. doi: 10.1086/340847
[44]
Nakamura T, Gulick J, Colbert MC, Robbins J (2009) Protein tyrosine phosphatase activity in the neural crest is essential for normal heart and skull development. Proc Natl Acad Sci U S A 106: 11270–11275. doi: 10.1073/pnas.0902230106
[45]
Nakamura T, Gulick J, Pratt R, Robbins J (2009) Noonan syndrome is associated with enhanced pERK activity, the repression of which can prevent craniofacial malformations. Proc Natl Acad Sci U S A 106: 15436–15441. doi: 10.1073/pnas.0903302106
[46]
Lapinski PE, Meyer MF, Feng GS, Kamiya N, King PD (2013) Deletion of SHP-2 in mesenchymal stem cells causes growth retardation, limb and chest deformity, and calvarial defects in mice. Dis Model Mech 6: 1448–1458. doi: 10.1242/dmm.012849
[47]
Bauler TJ, Kamiya N, Lapinski PE, Langewisch E, Mishina Y, et al. (2011) Development of severe skeletal defects in induced SHP-2-deficient adult mice: a model of skeletal malformation in humans with SHP-2 mutations. Dis Model Mech 4: 228–239.
