A Noncoding Point Mutation of Causes Multiple Developmental Malformations and Obesity in Twirler Mice

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Heterozygous Twirler (Tw) mice develop obesity and circling behavior associated with malformations of the inner ear, whereas homozygous Tw mice have cleft palate and die shortly after birth. Zeb1 is a zinc finger protein that contributes to mesenchymal cell fate by repression of genes whose expression defines epithelial cell identity. This developmental pathway is disrupted in inner ears of Tw/Tw mice. The purpose of our study was to comprehensively characterize the Twirler phenotype and to identify the causative mutation. The Tw/+ inner ear phenotype includes irregularities of the semicircular canals, abnormal utricular otoconia, a shortened cochlear duct, and hearing loss, whereas Tw/Tw ears are severely malformed with barely recognizable anatomy. Tw/+ mice have obesity associated with insulin-resistance and have lymphoid organ hypoplasia. We identified a noncoding nucleotide substitution, c.58+181G>A, in the first intron of the Tw allele of Zeb1 (Zeb1^Tw). A knockin mouse model of c.58+181G>A recapitulated the Tw phenotype, whereas a wild-type knockin control did not, confirming the mutation as pathogenic. c.58+181G>A does not affect splicing but disrupts a predicted site for Myb protein binding, which we confirmed in vitro. In comparison, homozygosity for a targeted deletion of exon 1 of mouse Zeb1, Zeb1^ΔEx1, is associated with a subtle abnormality of the lateral semicircular canal that is different than those in Tw mice. Expression analyses of E13.5 Twirler and Zeb1^ΔEx1 ears confirm that Zeb1^ΔEx1 is a null allele, whereas Zeb1^Tw RNA is expressed at increased levels in comparison to wild-type Zeb1. We conclude that a noncoding point mutation of Zeb1 acts via a gain-of-function to disrupt regulation of Zeb1^Tw expression, epithelial-mesenchymal cell fate or interactions, and structural development of the inner ear in Twirler mice. This is a novel mechanism underlying disorders of hearing or balance.

Vyšlo v časopise: A Noncoding Point Mutation of Causes Multiple Developmental Malformations and Obesity in Twirler Mice. PLoS Genet 7(9): e32767. doi:10.1371/journal.pgen.1002307
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1002307

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Zdroje

1. LyonMF 1958 Twirler: a mutant affecting the inner ear of the house mouse. J Embryol Exp Morphol 6 105 116

2. LanePWSearleAGBeecheyCVEicherE 1981 Chromosome 18 of the house mouse. J Hered 72 409 412

3. TingCNKohrmanDBurgessDLBoyleAAltschulerRA 1994 Insertional mutation on mouse chromosome 18 with vestibular and craniofacial abnormalities. Genetics 136 247 254

4. LiuHLiuWMaltbyKMLanYJiangR 2006 Identification and developmental expression analysis of a novel homeobox gene closely linked to the mouse Twirler mutation. Gene Expr Patterns 6 632 636

5. SekidoRMuraiKFunahashiJKamachiYFujisawa-SeharaA 1994 The delta-crystallin enhancer-binding protein delta EF1 is a repressor of E2-box-mediated gene activation. Mol Cell Biol 14 5692 5700

6. PostigoAADeanDC 1997 ZEB, a vertebrate homolog of Drosophila Zfh-1, is a negative regulator of muscle differentiation. EMBO J 16 3935 3943

7. ChamberlainEMSandersMM 1999 Identification of the novel player deltaEF1 in estrogen transcriptional cascades. Mol Cell Biol 19 3600 3606

8. PostigoAA 2003 Opposing functions of ZEB proteins in the regulation of the TGFbeta/BMP signaling pathway. EMBO J 22 2443 2452

9. NishimuraGManabeITsushimaKFujiuKOishiY 2006 DeltaEF1 mediates TGF-beta signaling in vascular smooth muscle cell differentiation. Dev Cell 11 93 104

10. TakagiTMoribeHKondohHHigashiY 1998 DeltaEF1, a zinc finger and homeodomain transcription factor, is required for skeleton patterning in multiple lineages. Development 125 21 31

11. SaykallyJNDoganSClearyMPSandersMM 2009 The ZEB1 transcription factor is a novel repressor of adiposity in female mice. PLoS One 4 e8460

12. PeinadoHOlmedaDCanoA 2007 Snail, Zeb and bHLH factors in tumour progression: an alliance against the epithelial phenotype? Nat Rev Cancer 7 415 428

13. HertzanoRElkonRKurimaKMorrissonAChanS-L 2011 Zeb1 and miR-200b regulate mesenchymal and epithelial cell identities in the developing mouse inner ear. Manuscript in preparation

14. KrafchakCMPawarHMoroiSESugarALichterPR 2005 Mutations in TCF8 cause posterior polymorphous corneal dystrophy and ectopic expression of COL4A3 by corneal endothelial cells. Am J Hum Genet 77 694 708

15. LiuYPengXTanJDarlingDSKaplanHJ 2008 Zeb1 mutant mice as a model of posterior corneal dystrophy. Invest Ophthalmol Vis Sci 49 1843 1849

16. HoweKMWatsonRJ 1991 Nucleotide preferences in sequence-specific recognition of DNA by c-myb protein. Nucleic Acids Res 19 3913 3919

17. KennedyAJEllacottKLKingVLHastyAH 2010 Mouse models of the metabolic syndrome. Dis Model Mech 3 156 166

18. ZhongZWanBQiuYNiJTangW 2007 Identification of a novel human zinc finger gene, ZNF438, with transcription inhibition activity. J Biochem Mol Biol 40 517 524

19. ChaykaOKintscherJBraasDKlempnauerKH 2005 v-Myb mediates cooperation of a cell-specific enhancer with the mim-1 promoter. Mol Cell Biol 25 499 511

20. KalueffAVIshikawaKGriffithAJ 2008 Anxiety and otovestibular disorders: linking behavioral phenotypes in men and mice. Behav Brain Res 186 1 11

21. CalderonADerrAStagnerBBJohnsonKRMartinG 2006 Cochlear developmental defect and background-dependent hearing thresholds in the Jackson circler (jc) mutant mouse. Hear Res 221 44 58

22. Stenzel-PooreMPCameronVAVaughanJSawchenkoPEValeW 1992 Development of Cushing's syndrome in corticotropin-releasing factor transgenic mice. Endocrinology 130 3378 3386

23. ShinaharaMNishiyamaMIwasakiYNakayamaSNoguchiT 2009 Plasma adiponectin levels are increased despite insulin resistance in corticotropin-releasing hormone transgenic mice, an animal model of Cushing syndrome. Endocr J 56 879 886

24. WangJScullyKZhuXCaiLZhangJ 2007 Opposing LSD1 complexes function in developmental gene activation and repression programmes. Nature 446 882 887

25. HagerJDinaCFranckeSDuboisSHouariM 1998 A genome-wide scan for human obesity genes reveals a major susceptibility locus on chromosome 10. Nat Genet 20 304 308

26. HinneyAZieglerAOeffnerFWedewardtCVogelM 2000 Independent confirmation of a major locus for obesity on chromosome 10. J Clin Endocrinol Metab 85 2962 2965

27. HsuehWCMitchellBDSchneiderJLSt JeanPLPollinTI 2001 Genome-wide scan of obesity in the Old Order Amish. J Clin Endocrinol Metab 86 1199 1205

28. PriceRALiWDBernsteinACrystalAGoldingEM 2001 A locus affecting obesity in human chromosome region 10p12. Diabetologia 44 363 366

29. CrynsKvan AlphenAMvan SpaendonckMPvan de HeyningPHTimmermansJP 2004 Circling behavior in the Ecl mouse is caused by lateral semicircular canal defects. J Comp Neurol 468 587 595

30. AdamsMEHurdEABeyerLASwiderskiDLRaphaelY 2007 Defects in vestibular sensory epithelia and innervation in mice with loss of Chd7 function: implications for human CHARGE syndrome. J Comp Neurol 504 519 532

31. BajpaiRChenDARada-IglesiasAZhangJXiongY 2010 CHD7 cooperates with PBAF to control multipotent neural crest formation. Nature 463 958 962

32. TellierALCormier-DaireVAbadieVAmielJSigaudyS 1998 CHARGE syndrome: report of 47 cases and review. Am J Med Genet 76 402 409

33. JongmansMCAdmiraalRJvan der DonkKPVissersLEBaasAF 2006 CHARGE syndrome: the phenotypic spectrum of mutations in the CHD7 gene. J Med Genet 43 306 314

34. ChangWLinZKulessaHHebertJHoganBL 2008 Bmp4 is essential for the formation of the vestibular apparatus that detects angular head movements. PLoS Genet 4 e1000050

35. VervoortRCeulemansHVan AerschotLD'HoogeRDavidG 2010 Genetic modification of the inner ear lateral semicircular canal phenotype of the Bmp4 haplo-insufficient mouse. Biochem Biophys Res Commun 394 780 785

36. MiyazonoKMaedaSImamuraT 2005 BMP receptor signaling: transcriptional targets, regulation of signals, and signaling cross-talk. Cytokine Growth Factor Rev 16 251 263

37. BeltranMPuigIPenaCGarciaJMAlvarezAB 2008 A natural antisense transcript regulates Zeb2/Sip1 gene expression during Snail1-induced epithelial-mesenchymal transition. Genes Dev 22 756 769

38. MizuguchiGKanei-IshiiCTakahashiTYasukawaTNagaseT 1995 c-Myb repression of c-erbB-2 transcription by direct binding to the c-erbB-2 promoter. J Biol Chem 270 9384 9389

39. TannoBSestiFCesiVBossiGFerrari-AmorottiG 2010 Expression of Slug is regulated by c-Myb and is required for invasion and bone marrow homing of cancer cells of different origin. J Biol Chem 285 29434 29445

40. DavisNYoffeCRavivSAntesRBergerJ 2009 Pax6 dosage requirements in iris and ciliary body differentiation. Dev Biol 333 132 142

41. FunkeBEpsteinJAKochilasLKLuMMPanditaRK 2001 Mice overexpressing genes from the 22q11 region deleted in velo-cardio-facial syndrome/DiGeorge syndrome have middle and inner ear defects. Hum Mol Genet 10 2549 2556

42. VitelliFViolaAMorishimaMPramparoTBaldiniA 2003 TBX1 is required for inner ear morphogenesis. Hum Mol Genet 12 2041 2048

43. VandewalleCVan RoyFBerxG 2009 The role of the ZEB family of transcription factors in development and disease. Cell Mol Life Sci 66 773 787

44. MountfordPZevnikBDuwelANicholsJLiM 1994 Dicistronic targeting constructs: reporters and modifiers of mammalian gene expression. Proc Natl Acad Sci U S A 91 4303 4307

45. TybulewiczVLCrawfordCEJacksonPKBronsonRTMulliganRC 1991 Neonatal lethality and lymphopenia in mice with a homozygous disruption of the c-abl proto-oncogene. Cell 65 1153 1163

46. KontgenFSussGStewartCSteinmetzMBluethmannH 1993 Targeted disruption of the MHC class II Aa gene in C57BL/6 mice. Int Immunol 5 957 964

47. HughesEDQuYYGenikSJLyonsRHPachecoCD 2007 Genetic variation in C57BL/6 ES cell lines and genetic instability in the Bruce4 C57BL/6 ES cell line. Mamm Genome 18 549 558

48. Ben-YosefTBelyantsevaIASaundersTLHughesEDKawamotoK 2003 Claudin 14 knockout mice, a model for autosomal recessive deafness DFNB29, are deaf due to cochlear hair cell degeneration. Hum Mol Genet 12 2049 2061

49. MorsliHChooDRyanAJohnsonRWuDK 1998 Development of the mouse inner ear and origin of its sensory organs. J Neurosci 18 3327 3335

50. BokJDolsonDKHillPRutherUEpsteinDJ 2007 Opposing gradients of Gli repressor and activators mediate Shh signaling along the dorsoventral axis of the inner ear. Development 134 1713 1722

51. NoguchiYKurimaKMakishimaTde AngelisMHFuchsH 2006 Multiple quantitative trait loci modify cochlear hair cell degeneration in the Beethoven (Tmc1Bth) mouse model of progressive hearing loss DFNA36. Genetics 173 2111 2119

52. KimHPennisiPAGavrilovaOPackSJouW 2006 Effect of adipocyte beta3-adrenergic receptor activation on the type 2 diabetic MKR mice. Am J Physiol Endocrinol Metab 290 E1227 1236

53. BorkJMPetersLMRiazuddinSBernsteinSLAhmedZM 2001 Usher syndrome 1D and nonsyndromic autosomal recessive deafness DFNB12 are caused by allelic mutations of the novel cadherin-like gene CDH23. Am J Hum Genet 68 26 37