Advertisement

Mammalian Genome

, Volume 15, Issue 6, pp 424–432 | Cite as

An ENU-induced mutation in AP-2α leads to middle earand ocular defects in Doarad mice

  • Nadav Ahituv
  • Alexandra Erven
  • Helmut Fuchs
  • Keren Guy
  • Ruth Ashery–Padan
  • Trevor Williams
  • Martin Hrabe de Angelis
  • Karen B. Avraham
  • Karen P. Steel
Article

Abstract

One of the advantages of N-ethyl- N-nitrosourea (ENU)-induced mutagenesis is that, after randomly causing point mutations, a variety of alleles can be generated in genes leading to diverse phenotypes. For example, transcription factor AP-2α (Tcfap2a) null homozygote mice show a large spectrum of developmental defects, among them missing middle ear bones and tympanic ring. This is the usual occurrence, where mutations causing middle ear anomalies usually coincide with other abnormalities. Using ENU-induced mutagenesis, we discovered a new dominant Tcfap2a mutant named Doarad (Dor) that has a missense mutation in the PY motif of its transactivation domain, leading to a misshapen malleus, incus, and stapes without any other observable phenotype. Dor homozygous mice die perinatally, showing prominent abnormal facial structures and ocular defects. In vitro assays suggest that this mutation causes a “gain of function” in the transcriptional activation of AP-2α. These mice enable us to address more specifically the developmental role of Tcfap2a in the eye and middle ear and are the first report of a mutation in a gene specifically causing middle ear abnormalities, leading to conductive hearing loss.

Keywords

Compound Action Potential Otosclerosis Endocochlear Potential Msx2 Compound Action Potential Amplitude 

Notes

Acknowledgments

The authors thank Helen Hurst, Stephanie Donaldson, Hubert Schorle, Thomas Schilling, and Ori Brenner for reagents and advice. We would like to acknowledge Rudi Balling for his contribution to setting up the mutagenesis program. This work was supported by the Israel Ministry of Health (K.B.A.), the G.I.F., the German-Israeli Foundation for Scientific Research and Development (K.B.A. and M.H.d.A.), the European Economic Community (QLG2-CT-1999-00988 and BMH4-CT97-2715; K.B.A. and K.P.S.), the UK MRC and Defeating Deafness (K.P.S.), the Israel Science Foundation (401/02; R.A.-P.), and NIH grant DE12728 (T.W.).

References

  1. Ashery–Padan, R, Marquardt, T, Zhou, X, Gruss, P 2000Pax6 activity in the lens primordium is required for lens formation and for correct placement of a single retina in the eyeGenes Dev1427012711CrossRefGoogle Scholar
  2. Bamforth, SD, Braganca, J, Eloranta, JJ, Murdoch, JN, Marques, FI,  et al. 2001Cardiac malformations, adrenal agenesis, neural crest defects and exencephaly in mice lacking Cited2, a new Tfap2 co-activatorNat Genet29469474CrossRefGoogle Scholar
  3. Bosher, JM, Totty, NF, Hsuan, JJ, Williams, T, Hurst, HC 1996A family of AP-2 proteins regulates c-erbB-2 expression in mammary carcinomaOncogene1317011707PubMedGoogle Scholar
  4. Braganca, J, Swingler, T, Marques, FI, Jones, T, Eloranta, JJ,  et al. 2002Human CREB-binding protein/p300-interacting transactivator with ED-rich tail (CITED) 4, a new member of the CITED family, functions as a co-activator for transcription factor AP-2J Biol Chem27785598565CrossRefGoogle Scholar
  5. Browning, GG, Gatehouse, S 1989Hearing in chronic suppurative otitis mediaAnn Oto Rhinol Laryngol98245250CrossRefGoogle Scholar
  6. Chen, HI, Sudol, M 1995The WW domain of Yes-associated protein binds a proline-rich ligand that differs from the consensus established for Src homology 3-binding modulesProc Natl Acad Sci USA9278197823CrossRefGoogle Scholar
  7. Declau, F, Heyning, P 1996OtosclerosisMartini, ARead, AStephens, D eds. Genetics and Hearing Impairment,WhurrLondon, UK221230Google Scholar
  8. Evans, EF 1975The sharpening of cochlear frequency selectivity in the normal and abnormal cochleaAudiology14419442CrossRefGoogle Scholar
  9. Hermanns, P, Lee, B 2001Transcriptional dysregulation in skeletal malformation syndromesAm J Med Genet106258271CrossRefGoogle Scholar
  10. Hilger–Eversheim, K, Moser, M, Schorle, H, Buettner, R 2000Regulatory roles of AP-2 transcription factors in vertebrate development, apoptosis and cell-cycle controlGene260112CrossRefGoogle Scholar
  11. de Hrabe Angelis, MH, Flaswinkel, H, Fuchs, H, Rathkolb, B, Soewarto, D,  et al. 2000Genome- wide, large-scale production of mutant mice by ENU mutagenesisNat Genet25444447CrossRefGoogle Scholar
  12. Ilsley, JL, Sudol, M, Winder, SJ 2002The WW domain: linking cell signalling to the membrane cytoskeletonCell Signal14183189CrossRefGoogle Scholar
  13. Kaiser, K, Stelzer, G, Meisterernst, M 1995The coactivator p15 (PC4) initiates transcriptional activation during TFIIA–TFIID-promoter complex formationEMBO J1435203527PubMedPubMedCentralGoogle Scholar
  14. Kannan, P, Tainsky, MA 1999Coactivator PC4 mediates AP-2 transcriptional activity and suppresses ras-induced transformation dependent on AP-2 transcriptional interferenceMol Cell Biol19899908CrossRefGoogle Scholar
  15. Kiernan, AE, Zalzman, M, Fuchs, H, Hrabe de Angelis, MH, Balling, R,  et al. 1999Tailchaser (Tlc): a new mouse mutation affecting hair bundle differentiation and hair cell survivalJ Neurocytol28969985CrossRefGoogle Scholar
  16. Kiernan, AE, Erven, A, Voegeling, S, Peters, J, Nolan, P,  et al. 2002ENU mutagenesis reveals a highly mutable locus on mouse Chromosome 4 that affects ear morphogenesisMamm Genome13142148CrossRefGoogle Scholar
  17. Knight, RD, Nair, S, Nelson, SS, Afshar, A, Javidan, Y,  et al. 2003Lockjaw encodes a zebrafish tfap2a required for early neural crest developmentDevelopment13057555768CrossRefGoogle Scholar
  18. Law, CJ, Fisher, AM, Temple, IK 1998Distal 6p deletion syndrome: a report of a case with anterior chamber eye anomaly and review of published reportsJ Med Genet35685689CrossRefGoogle Scholar
  19. Liu, YH, Kundu, R, Wu, L, Luo, W, Ignelzi Jr, MA,  et al. 1995Premature suture closure and ectopic cranial bone in mice expressing Msx2 transgenes in the developing skullProc Natl Acad Sci USA9261376141CrossRefGoogle Scholar
  20. Ma, L, Golden, S, Wu, L, Maxson, R 1996The molecular basis of Boston-type craniosynostosis: the Pro l48→His mutation in the N-terminal arm of the MSX2 homeodomain stabilizes DNA binding without altering nucleotide sequence preferencesHum Mol Genet519151920CrossRefGoogle Scholar
  21. Mallo, M 1998Embryological and genetic aspects of middle ear developmentInt J Dev Biol421122PubMedGoogle Scholar
  22. Mitchell, PJ, Timmons, PM, Hebert, JM, Rigby, PW, Tjian, R 1991Transcription factor AP-2 is expressed in neural crest cell lineages during mouse embryogenesisGenes Dev5105119CrossRefGoogle Scholar
  23. Nottoli, T, Hagopian–Donaldson, S, Zhang, J, Perkins, A, Williams, T 1998AP-2-null cells disrupt morphogenesis of the eye, face, and limbs in chimeric miceProc Natl Acad Sci USA951371413719CrossRefGoogle Scholar
  24. Pirozzi, G, McConnell, SJ, Uveges, AJ, Carter, JM, Sparks, AB,  et al. 1997Identification of novel human WW domain-containing proteins by cloning of ligand targetsJ Biol Chem2721461114616CrossRefGoogle Scholar
  25. Pogulis, RJ, Vallejo, AN, Pease, LR 1996In vitro recombination and mutagenesis by overlap extension PCRTrower, MK eds. In vitro mutagenesis protocolsHumana Press IncTotowa, NJ167176CrossRefGoogle Scholar
  26. Satokata, I, Ma, L, Ohshima, H, Bei, M, Woo, I,  et al. 2000Msx2 deficiency in mice causes pleiotropic defects in bone growth and ectodermal organ formationNat Genet24391395CrossRefGoogle Scholar
  27. Schorle, H, Meier, P, Buchert, M, Jaenisch, R, Mitchell, PJ 1996Transcription factor AP-2 essential for cranial closure and craniofacial developmentNature381235238CrossRefGoogle Scholar
  28. Steel, KP, Smith, RJ 1992Normal hearing in Splotch (Sp/+), the mouse homologue of Waardenburg syndrome type 1Nat Genet27579CrossRefGoogle Scholar
  29. Wankhade, S, Yu, Y, Weinberg, J, Tainsky, MA, Kannan, P 2000Characterization of the activation domains of AP-2 family transcription factorsJ Biol Chem2752970129708CrossRefGoogle Scholar
  30. West–Mays, JA, Zhang, J, Nottoli, T, Hagopian-Donaldson, S, Libby, D,  et al. 1999AP-2alpha transcription factor is required for early morphogenesis of the lens vesicleDev Biol2064662CrossRefGoogle Scholar
  31. Williams, T, Tjian, R 1991Analysis of the DNA-binding and activation properties of the human transcription factor AP-2Genes Dev5670682CrossRefGoogle Scholar
  32. Yagi, R, Chen, LF, Shigesada, K, Murakami, Y, Ito, Y 1999A WW domain-containing yes-associated protein (YAP) is a novel transcriptional co-activatorEMBO J1825512562CrossRefGoogle Scholar
  33. Zhang, J, Hagopian–Donaldson, S, Serbedzija, G, Elsemore, J, Plehn-Dujowich, D,  et al. 1996Neural tube, skeletal and body wall defects in mice lacking transcription factor AP-2Nature381238241CrossRefGoogle Scholar
  34. Zhao, F, Satoda, M, Licht, JD, Hayashizaki, Y, Gelb, BD 2001aCloning and characterization of a novel mouse AP-2 transcription factor, AP-2delta, with unique DNA binding and transactivation propertiesJ Biol Chem2764075540760CrossRefGoogle Scholar
  35. Zhao, F, Weismann, CG, Satoda, M, Pierpont, ME, Sweeney, E,  et al. 2001bNovel TFAP2B mutations that cause Char syndrome provide a genotype-phenotype correlationAm J Hum Genet69695703CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Nadav Ahituv
    • 1
  • Alexandra Erven
    • 2
  • Helmut Fuchs
    • 3
  • Keren Guy
    • 1
  • Ruth Ashery–Padan
    • 1
  • Trevor Williams
    • 4
  • Martin Hrabe de Angelis
    • 3
  • Karen B. Avraham
    • 1
  • Karen P. Steel
    • 2
  1. 1.Department of Human Genetics and Molecular Medicine, Sackler School of MedicineTel Aviv UniversityTel AvivIsrael
  2. 2.MRC Institute of Hearing ResearchUniversity ParkNottinghamUK
  3. 3.GSF Research Center for Environment and HealthInstitute of Experimental GeneticsNeuherbergGermany
  4. 4.Departments of Craniofacial Biology and of Cell and Structural BiologyUniversity of Colorado Health Sciences CenterDenverUSA

Personalised recommendations