Characterization of a novel missense mutation on murine Pax3 through ENU mutagenesis

Yue Xiao; Lingling Zhang; Kuanjun He; Xiang Gao; Lun Yang; Lin He; Gang Ma; Xizhi Guo

doi:10.1016/j.jgg.2011.06.005

Volume 38 Issue 8

Aug. 2011

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Characterization of a novel missense mutation on murine Pax3 through ENU mutagenesis

doi: 10.1016/j.jgg.2011.06.005

a
Bio-X Center, Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders, Shanghai Jiao Tong University, Shanghai 200240, China
b
Medical School of Nanjing University, Model Animal Research Center, Nanjing 210093, China

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Corresponding author: E-mail address: xzguo2005@sjtu.edu.cn (Xizhi Guo)
Received Date: 2011-04-01
Accepted Date: 2011-06-12
Rev Recd Date: 2011-06-07

Available Online: 2011-07-19

Publish Date: 2011-08-20

Abstract

Abstract

N-ethyl-N-nitrosourea (ENU) mutagenesis has led to the elucidation of several regulator genes for melanocyte and skin development. Here we characterized a mutant from ENU mutagenesis with similar phenotype as that of Splotch mutant, including exencephaly, spina bifida and abnormal limbs in homozygotes as well as white belly spotting and occasionally loop-tail in heterozygotes. This novel mutant was named as . Through genome-wide linkage analysis in backcross progenies with microsatellite markers, the was confined to a region between D1MIT415 and D1MIT7 on chromosome 1, where notable Pax3 gene was located. Direct sequencing revealed that carried a nucleotide A894G missense transition in exon 6 of Pax3 gene that resulted in Asn to Asp substitution at amino acid 269 within the highly-conserved homeodomain (HD) DNA recognition module, which was the first point mutation found in this domain in mice. This N269D mutation impaired the transactivation capacity of Pax3 protein, but exerted no effect on Pax3 protein translation. The characterization of the new mutation expanded our understanding the transactivation and DNA-binding structure of Pax3 protein.
- White spotting,
- Neural tube defect,
- ENU,
- Pax3,
- Transactivation

These authors contributed equally to this paper.

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References(25)

References

[1]	Apuzzo, S., Abdelhakim, A., Fortin, A.S. et al. Cross-talk between the paired domain and the homeodomain of Pax3: DNA binding by each domain causes a structural change in the other domain, supporting interdependence for DNA binding J. Biol. Chem., 279 (2004),pp. 33601-33612
[2]	Barr, F.G., Galili, N., Holick, J. et al. Nat. Genet., 3 (1993),pp. 113-117
[3]	Birrane, G., Soni, A., Ladias, J.A. Structural basis for DNA recognition by the human PAX3 homeodomain Biochemistry, 48 (2009),pp. 1148-1155
[4]	Conway, S.J., Henderson, D.J., Kirby, M.L. et al. Cardiovasc. Res., 36 (1997),pp. 163-173
[5]	Epstein, D.J., Vekemans, M., Gros, P. Splotch (Sp2H), a mutation affecting development of the mouse neural tube, shows a deletion within the paired homeodomain of Pax-3 Cell, 67 (1991),pp. 767-774
[6]	Epstein, D.J., Vogan, K.J., Trasler, D.G. et al. Proc. Natl. Acad. Sci. USA, 90 (1993),pp. 532-536
[7]	Farrer, L.A., Arnos, K.S., , Baldwin, C.T. et al. Locus heterogeneity for Waardenburg syndromeis predictive of clinical subtypes Am. J. Hum. Genet., 55 (1994),pp. 728-737
[8]	Glogarova, K., Buckiova, D. Changes in sialylation in homozygous Sp2H mouse mutant embryos Birth Defects Res. A Clin. Mol. Teratol., 70 (2004),pp. 142-152
[9]	Guo, X.L., Ruan, H.B., Li, Y. et al. Pigment Cell Melanoma Res., 23 (2010),pp. 252-262
[10]	Lam, P.Y., Sublett, J.E., Hollenbach, A.D. et al. The oncogenic potential of the Pax3-FKHR fusion protein requires the Pax3 homeodomain recognition helix but not the Pax3 paired-box DNA binding domain Mol. Cell. Biol., 19 (1999),pp. 594-601
[11]	Lang, D., Epstein, J.A. Sox10 and Pax3 physically interact to mediate activation of a conserved c-RET enhancer Hum. Mol. Genet., 12 (2003),pp. 937-945
[12]	Lang, D., Lu, M.M., Huang, L. et al. Pax3 functions at a nodal point in melanocyte stem cell differentiation Nature, 433 (2005),pp. 884-887
[13]	Machado, A.F., Martin, L.J., Collins, M.D. Pax3 and the splotch mutations: structure, function, and relationship to teratogenesis, including gene-chemical interactions Curr. Pharm. Des., 7 (2001),pp. 751-785
[14]	Milewski, R.C., Chi, N.C., Li, J. et al. Identification of minimal enhancer elements sufficient for Pax3 expression in neural crest and implication of Tead2 as a regulator of Pax3 Development, 131 (2004),pp. 829-837
[15]	Pasteris, N.G., Trask, B.J., Sheldon, S. et al. Hum. Mol. Genet., 2 (1993),pp. 953-959
[16]	Phelan, S.A., Loeken, M.R. Identification of a new binding motif for the paired domain of Pax-3 and unusual characteristics of spacing of bipartite recognition elements on binding and transcription activation J. Biol. Chem., 273 (1998),pp. 19153-19159
[17]	Pingault, V., Ente, D., Dastot-Le Moal, F. et al. Review and update of mutations causing Waardenburg syndrome Hum. Genet., 31 (2010),pp. 391-406
[18]	Potterf, S.B., Furumura, M., Dunn, K.J. et al. Transcription factor hierarchy in Waardenburg syndrome: regulation of MITF expression by SOX10 and PAX3 Hum. Genet., 107 (2000),pp. 1-6
[19]	Pritchard, C., Grosveld, G., Hollenbach, A.D. Gene, 305 (2003),pp. 61-69
[20]	Relaix, F., Rocancourt, D., Mansouri, A. et al. Divergent functions of murine Pax3 and Pax7 in limb muscle development Genes Dev., 18 (2004),pp. 1088-1105
[21]	Russell, B. Some possibilities in the prevention of disorders of the skin J. R. Inst. Public Health, 10 (1947),pp. 191-205
[22]	Steingrimsson, E., Copeland, N.G., Jenkins, N.A. Melanocyte stem cell maintenance and hair graying Cell, 121 (2005),pp. 9-12
[23]	Tassabehji, M., Read, A.P., Newild-Typeon, V.E. et al. Nat. Genet., 3 (1993),pp. 26-30
[24]	Watanabe, A., Takeda, K., Ploplis, B. et al. Epistatic relationship between Waardenburg syndrome genes MITF and PAX3 Nat. Genet., 18 (1998),pp. 283-286
[25]	Wu, M., Li, J., Engleka, K.A. et al. J. Clin. Invest., 118 (2008),pp. 2076-2087