Use of a conditional ubr5 mutant allele to investigate the role of an n-End rule ubiquitin-protein ligase in hedgehog signalling and embryonic limb development

PLoS ONE

Volumn 11, Issue 6, 2016, Pages

  Kinsella, Elaine ^a   Dora, Natalie ^b   Mellis, David ^a   Lettice, Laura ^b   Deveney, Paul ^b   Hill, Robert ^b   Ditzel, Mark ^a  

^a UNIVERSITY OF EDINBURGH   (United Kingdom)

^b UNIVERSITY OF EDINBURGH   (United Kingdom)

Author keywords

[No Author keywords available]

Indexed keywords

SHORT HAIRPIN RNA; SONIC HEDGEHOG PROTEIN; UBIQUITIN PROTEIN LIGASE; RETINOIC ACID; UBR5 PROTEIN, MOUSE;

ADULT; ANIMAL CELL; ARTICLE; CONTROLLED STUDY; EMBRYO; EMBRYONIC STEM CELL; FEMALE; GENE; GENE EXPRESSION; GENE FREQUENCY; IN VIVO STUDY; LIMB DEVELOPMENT; LOSS OF FUNCTION MUTATION; MALE; MOUSE; NONHUMAN; RNA INTERFERENCE; SHH GENE; SIGNAL TRANSDUCTION; UBR5 GENE; ABNORMALITIES; ALLELE; ANIMAL; CELL LINE; CYTOLOGY; EMBRYOLOGY; GENE DELETION; GENE EXPRESSION REGULATION; GENETICS; LIMB; LIMB BUD; LIMB MALFORMATION; METABOLISM; MOUSE EMBRYONIC STEM CELL; MUTATION; PHENOTYPE; PHYSIOLOGY;

ALLELES; ANIMALS; CELL LINE; EXTREMITIES; GENE DELETION; GENE EXPRESSION REGULATION, DEVELOPMENTAL; HEDGEHOG PROTEINS; LIMB BUDS; LIMB DEFORMITIES, CONGENITAL; MICE; MOUSE EMBRYONIC STEM CELLS; MUTATION; PHENOTYPE; SIGNAL TRANSDUCTION; TRETINOIN; UBIQUITIN-PROTEIN LIGASES;

EID: 84976407367     PISSN: None     EISSN: 19326203     Source Type: Journal    
DOI: 10.1371/journal.pone.0157079     Document Type: Article

References (77)

1. Briscoe J, Therond PP. The mechanisms of Hedgehog signalling and its roles in development and disease. Nature reviews Molecular cell biology. 2013; 14(7):416-29. Epub 2013/05/31. doi: 10.1038/ nrm3598 PMID: 23719536.
2. Petrova R, Joyner AL. Roles for Hedgehog signaling in adult organ homeostasis and repair. Development. 2014; 141(18):3445-57. doi: 10.1242/dev.083691 PMID: 25183867; PubMed Central PMCID: PMCPMC4197719.
3. Towers M, Wolpert L, Tickle C. Gradients of signalling in the developing limb. Curr Opin Cell Biol. 2012; 24(2):181-7. doi: 10.1016/j.ceb.2011.11.005 PMID: 22169676.
4. Takebe N, Miele L, Harris PJ, Jeong W, Bando H, Kahn M, et al. Targeting Notch, Hedgehog, and Wnt pathways in cancer stem cells: clinical update. Nat Rev Clin Oncol. 2015; 12(8):445-64. doi: 10.1038/ nrclinonc.2015.61 PMID: 25850553; PubMed Central PMCID: PMCPMC4520755.
5. McMahon AP, Ingham PW, Tabin CJ. Developmental roles and clinical significance of hedgehog signaling. Curr Top Dev Biol. 2003; 53:1-114. PMID: 12509125.
6. Chen Y, Struhl G. Dual roles for patched in sequestering and transducing Hedgehog. Cell. 1996; 87 (3):553-63. PMID: 8898207.
7. Marigo V, Davey RA, Zuo Y, Cunningham JM, Tabin CJ. Biochemical evidence that patched is the Hedgehog receptor. Nature. 1996; 384(6605):176-9. Epub 1996/11/14. doi: 10.1038/384176a0 PMID: 8906794.
8. Stone DM, Hynes M, Armanini M, Swanson TA, Gu Q, Johnson RL, et al. The tumour-suppressor gene patched encodes a candidate receptor for Sonic hedgehog. Nature. 1996; 384(6605):129-34. Epub 1996/11/14. doi: 10.1038/384129a0 PMID: 8906787.
9. Towers M, Tickle C. Growing models of vertebrate limb development. Development. 2009; 136(2):179-90. doi: 10.1242/dev.024158 PMID: 19103802.
10. Riddle RD, Johnson RL, Laufer E, Tabin C. Sonic hedgehog mediates the polarizing activity of the ZPA. Cell. 1993; 75(7):1401-16. PMID: 8269518.
11. Lettice LA, Heaney SJ, Purdie LA, Li L, de Beer P, Oostra BA, et al. A long-range Shh enhancer regulates expression in the developing limb and fin and is associated with preaxial polydactyly. Hum Mol Genet. 2003; 12(14):1725-35. PMID: 12837695.
12. Hill RE, Lettice LA. Alterations to the remote control of Shh gene expression cause congenital abnormalities. Philos Trans R Soc Lond B Biol Sci. 2013; 368(1620):20120357. doi: 10.1098/rstb.2012.0357 PMID: 23650631; PubMed Central PMCID: PMCPMC3682722.
13. Anderson E, Hill RE. Long range regulation of the sonic hedgehog gene. Curr Opin Genet Dev. 2014; 27:54-9. doi: 10.1016/j.gde.2014.03.011 PMID: 24859115.
14. Lauth M, Bergstrom A, Shimokawa T, Tostar U, Jin Q, Fendrich V, et al. DYRK1B-dependent autocrine-to-paracrine shift of Hedgehog signaling by mutant RAS. Nat Struct Mol Biol. 2010; 17(6):718-25. doi: 10.1038/nsmb.1833 PMID: 20512148.
15. Lee JD, Amanai K, Shearn A, Treisman JE. The ubiquitin ligase Hyperplastic discs negatively regulates hedgehog and decapentaplegic expression by independent mechanisms. Development. 2002; 129 (24):5697-706. PMID: 12421709.
16. Moncrieff S, Moncan M, Scialpi F, Ditzel M. Regulation of hedgehog Ligand Expression by the N-End Rule Ubiquitin-Protein Ligase Hyperplastic Discs and the Drosophila GSK3beta Homologue, Shaggy. PLoS One. 2015; 10(9):e0136760. doi: 10.1371/journal.pone.0136760 PMID: 26334301; PubMed Central PMCID: PMCPMC4559392.
17. Kozlov G, Nguyen L, Lin T, De Crescenzo G, Park M, Gehring K. Structural basis of ubiquitin recognition by the ubiquitin-associated (UBA) domain of the ubiquitin ligase EDD. J Biol Chem. 2007; 282 (49):35787-95. PMID: 17897937.
18. Matta-Camacho E, Kozlov G, Li FF, Gehring K. Structural basis of substrate recognition and specificity in the N-end rule pathway. Nat Struct Mol Biol. 2010; 17(10):1182-7. Epub 2010/09/14. doi: 10.1038/ nsmb.1894 PMID: 20835242.
19. Tasaki T, Mulder LC, Iwamatsu A, Lee MJ, Davydov IV, Varshavsky A, et al. A family of mammalian E3 ubiquitin ligases that contain the UBR box motif and recognize N-degrons. Mol Cell Biol. 2005; 25 (16):7120-36. PMID: 16055722.
20. Matta-Camacho E, Kozlov G, Menade M, Gehring K. Structure of the HECT C-lobe of the UBR5 E3 ubiquitin ligase. Acta Crystallogr Sect F Struct Biol Cryst Commun. 2012; 68(Pt 10):1158-63. Epub 2012/ 10/03. doi: 10.1107/S1744309112036937 PMID: 23027739; PubMed Central PMCID: PMC3497971.
21. Gudjonsson T, Altmeyer M, Savic V, Toledo L, Dinant C, Grofte M, et al. TRIP12 and UBR5 suppress spreading of chromatin ubiquitylation at damaged chromosomes. Cell. 2012; 150(4):697-709. Epub 2012/08/14. doi: 10.1016/j.cell.2012.06.039 PMID: 22884692.
22. Okamoto K, Bartocci C, Ouzounov I, Diedrich JK, Yates JR 3rd, Denchi EL. A two-step mechanism for TRF2-mediated chromosome-end protection. Nature. 2013; 494(7438):502-5. Epub 2013/02/08. doi: 10.1038/nature11873 PMID: 23389450; PubMed Central PMCID: PMC3733551.
23. Zhang T, Cronshaw J, Kanu N, Snijders AP, Behrens A. UBR5-mediated ubiquitination of ATMIN is required for ionizing radiation-induced ATM signaling and function. Proceedings of the National Academy of Sciences of the United States of America. 2014; 111(33):12091-6. Epub 2014/08/06. doi: 10. 1073/pnas.1400230111 PMID: 25092319; PubMed Central PMCID: PMC4143036.
24. Su H, Meng S, Lu Y, Trombly MI, Chen J, Lin C, et al. Mammalian hyperplastic discs homolog EDD regulates miRNA-mediated gene silencing. Molecular cell. 2011; 43(1):97-109. Epub 2011/07/06. doi: 10. 1016/j.molcel.2011.06.013 PMID: 21726813; PubMed Central PMCID: PMC3147021.
25. Reid MA, Wang WI, Rosales KR, Welliver MX, Pan M, Kong M. The B55alpha subunit of PP2A drives a p53-dependent metabolic adaptation to glutamine deprivation. Molecular cell. 2013; 50(2):200-11. Epub 2013/03/19. doi: 10.1016/j.molcel.2013.02.008 PMID: 23499005.
26. Benavides M, Chow-Tsang LF, Zhang J, Zhong H. The novel interaction between microspherule protein Msp58 and ubiquitin E3 ligase EDD regulates cell cycle progression. Biochimica et biophysica acta. 2013; 1833(1):21-32. Epub 2012/10/17. doi: 10.1016/j.bbamcr.2012.10.007 PMID: 23069210; PubMed Central PMCID: PMC3518595.
27. Maddika S, Chen J. Protein kinase DYRK2 is a scaffold that facilitates assembly of an E3 ligase. Nat Cell Biol. 2009; 11(4):409-19. PMID: 19287380. doi: 10.1038/ncb1848
28. Munoz MA, Saunders DN, Henderson MJ, Clancy JL, Russell AJ, Lehrbach G, et al. The E3 ubiquitin ligase EDD regulates S-phase and G(2)/M DNA damage checkpoints. Cell cycle. 2007; 6(24):3070-7. Epub 2007/12/13. PMID: 18073532.
29. Scialpi F, Mellis D, Ditzel M. EDD, a ubiquitin-protein ligase of the N-end rule pathway, associates with spindle assembly checkpoint components and regulates the mitotic response to nocodazole. The Journal of biological chemistry. 2015; 290(20):12585-94. Epub 2015/04/03. doi: 10.1074/jbc.M114.625673 PMID: 25833949; PubMed Central PMCID: PMC4432279.
30. Smits VA. EDD induces cell cycle arrest by increasing p53 levels. Cell cycle. 2012; 11(4). Epub 2012/ 02/07. PMID: 22306983.
31. Yuan JS, Reed A, Chen F, Stewart CN Jr. Statistical analysis of real-time PCR data. BMC Bioinformatics. 2006; 7:85. doi: 10.1186/1471-2105-7-85 PMID: 16504059; PubMed Central PMCID: PMCPMC1395339.
32. Ditzel M, Broemer M, Tenev T, Bolduc C, Lee TV, Rigbolt KT, et al. Inactivation of effector caspases through nondegradative polyubiquitylation. Mol Cell. 2008; 32(4):540-53. PMID: 19026784. doi: 10. 1016/j.molcel.2008.09.025
33. Truett GE, Heeger P, Mynatt RL, Truett AA, Walker JA, Warman ML. Preparation of PCR-quality mouse genomic DNA with hot sodium hydroxide and tris (HotSHOT). Biotechniques. 2000; 29(1):52, 4. PMID: 10907076.
34. Buchholz F, Angrand PO, Stewart AF. Improved properties of FLP recombinase evolved by cycling mutagenesis. Nat Biotechnol. 1998; 16(7):657-62. doi: 10.1038/nbt0798-657 PMID: 9661200.
35. Indra AK, Warot X, Brocard J, Bornert JM, Xiao JH, Chambon P, et al. Temporally-controlled site-specific mutagenesis in the basal layer of the epidermis: comparison of the recombinase activity of the tamoxifen-inducible Cre-ER(T) and Cre-ER(T2) recombinases. Nucleic Acids Res. 1999; 27(22):4324-7. PMID: 10536138; PubMed Central PMCID: PMCPMC148712.
36. Logan M, Martin JF, Nagy A, Lobe C, Olson EN, Tabin CJ. Expression of Cre Recombinase in the developing mouse limb bud driven by a Prxl enhancer. Genesis. 2002; 33(2):77-80. Epub 2002/07/12. doi: 10.1002/gene.10092 PMID: 12112875.
37. Sharpe J, Ahlgren U, Perry P, Hill B, Ross A, Hecksher-Sorensen J, et al. Optical projection tomography as a tool for 3D microscopy and gene expression studies. Science. 2002; 296(5567):541-5. doi: 10.1126/science.1068206 PMID: 11964482.
38. Okada Y, Shimazaki T, Sobue G, Okano H. Retinoic-acid-concentration-dependent acquisition of neural cell identity during in vitro differentiation of mouse embryonic stem cells. Dev Biol. 2004; 275 (1):124-42. doi: 10.1016/j.ydbio.2004.07.038 PMID: 15464577.
39. Williams RL, Hilton DJ, Pease S, Willson TA, Stewart CL, Gearing DP, et al. Myeloid leukaemia inhibitory factor maintains the developmental potential of embryonic stem cells. Nature. 1988; 336 (6200):684-7. doi: 10.1038/336684a0 PMID: 3143916.
40. Okabe S, Forsberg-Nilsson K, Spiro AC, Segal M, McKay RD. Development of neuronal precursor cells and functional postmitotic neurons from embryonic stem cells in vitro. Mech Dev. 1996; 59(1):89-102. PMID: 8892235.
41. Bain G, Kitchens D, Yao M, Huettner JE, Gottlieb DI. Embryonic stem cells express neuronal properties in vitro. Dev Biol. 1995; 168(2):342-57. doi: 10.1006/dbio.1995.1085 PMID: 7729574.
42. Chambers I, Colby D, Robertson M, Nichols J, Lee S, Tweedie S, et al. Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell. 2003; 113(5):643-55. PMID: 12787505.
43. Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 2006; 126(4):663-76. doi: 10.1016/j.cell.2006.07.024 PMID: 16904174.
44. Qi X, Li TG, Hao J, Hu J, Wang J, Simmons H, et al. BMP4 supports self-renewal of embryonic stem cells by inhibiting mitogen-activated protein kinase pathways. Proc Natl Acad Sci U S A. 2004; 101 (16):6027-32. doi: 10.1073/pnas.0401367101 PMID: 15075392; PubMed Central PMCID: PMCPMC395917.
45. Saunders DN, Hird SL, Withington SL, Dunwoodie SL, Henderson MJ, Biben C, et al. Edd, the murine hyperplastic disc gene, is essential for yolk sac vascularization and chorioallantoic fusion. Mol Cell Biol. 2004; 24(16):7225-34. PMID: 15282321.
46. Schnutgen F, De-Zolt S, Van Sloun P, Hollatz M, Floss T, Hansen J, et al. Genomewide production of multipurpose alleles for the functional analysis of the mouse genome. Proc Natl Acad Sci U S A. 2005; 102(20):7221-6. doi: 10.1073/pnas.0502273102 PMID: 15870191; PubMed Central PMCID: PMCPMC1129123.
47. Lim NS, Kozlov G, Chang TC, Groover O, Siddiqui N, Volpon L, et al. Comparative peptide binding studies of the PABC domains from the ubiquitin-protein isopeptide ligase HYD and poly(A)-binding protein. Implications for HYD function. J Biol Chem. 2006; 281(20):14376-82. PMID: 16554297.
48. Chiang C, Litingtung Y, Lee E, Young KE, Corden JL, Westphal H, et al. Cyclopia and defective axial patterning in mice lacking Sonic hedgehog gene function. Nature. 1996; 383(6599):407-13. PMID: 8837770.
49. Tsukui T, Capdevila J, Tamura K, Ruiz-Lozano P, Rodriguez-Esteban C, Yonei-Tamura S, et al. Multiple left-right asymmetry defects in Shh(-/-) mutant mice unveil a convergence of the shh and retinoic acid pathways in the control of Lefty-1. Proc Natl Acad Sci U S A. 1999; 96(20):11376-81. PMID: 10500184; PubMed Central PMCID: PMCPMC18041.
50. Feil R, Wagner J, Metzger D, Chambon P. Regulation of Cre recombinase activity by mutated estrogen receptor ligand-binding domains. Biochem Biophys Res Commun. 1997; 237(3):752-7. doi: 10.1006/ bbrc.1997.7124 PMID: 9299439.
51. Niwa H, Yamamura K, Miyazaki J. Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene. 1991; 108(2):193-9. PMID: 1660837.
52. Vortkamp A, Lee K, Lanske B, Segre GV, Kronenberg HM, Tabin CJ. Regulation of rate of cartilage differentiation by Indian hedgehog and PTH-related protein. Science. 1996; 273(5275):613-22. PMID: 8662546.
53. St-Jacques B, Hammerschmidt M, McMahon AP. Indian hedgehog signaling regulates proliferation and differentiation of chondrocytes and is essential for bone formation. Genes Dev. 1999; 13(16):2072-86.PMID: 10465785; PubMed Central PMCID: PMCPMC316949.
54. Visel A, Thaller C, Eichele G. GenePaint.org: an atlas of gene expression patterns in the mouse embryo. Nucleic Acids Res. 2004; 32(Database issue):D552-6. doi: 10.1093/nar/gkh029 PMID: 14681479; PubMed Central PMCID: PMCPMC308763.
55. Sears KE, Maier JA, Rivas-Astroza M, Poe R, Zhong S, Kosog K, et al. The Relationship between Gene Network Structure and Expression Variation among Individuals and Species. PLoS Genet. 2015; 11(8):e1005398. doi: 10.1371/journal.pgen.1005398 PMID: 26317994; PubMed Central PMCID: PMCPMC4552942.
56. Niedermaier M, Schwabe GC, Fees S, Helmrich A, Brieske N, Seemann P, et al. An inversion involving the mouse Shh locus results in brachydactyly through dysregulation of Shh expression. J Clin Invest. 2005; 115(4):900-9. doi: 10.1172/JCI23675 PMID: 15841179; PubMed Central PMCID: PMCPMC1070420.
57. Petiot A, Conti FJ, Grose R, Revest JM, Hodivala-Dilke KM, Dickson C. A crucial role for Fgfr2-IIIb signalling in epidermal development and hair follicle patterning. Development. 2003; 130(22):5493-501. doi: 10.1242/dev.00788 PMID: 14530295.
58. Henderson MJ, Russell AJ, Hird S, Munoz M, Clancy JL, Lehrbach GM, et al. EDD, the human hyperplastic discs protein, has a role in progesterone receptor coactivation and potential involvement in DNA damage response. J Biol Chem. 2002; 277(29):26468-78. PMID: 12011095.
59. Clarke CL, Roman SD, Graham J, Koga M, Sutherland RL. Progesterone receptor regulation by retinoic acid in the human breast cancer cell line T-47D. J Biol Chem. 1990; 265(21):12694-700. PMID: 2373707.
60. Matsumoto H, Zhao X, Das SK, Hogan BL, Dey SK. Indian hedgehog as a progesterone-responsive factor mediating epithelial-mesenchymal interactions in the mouse uterus. Dev Biol. 2002; 245(2):280-90. doi: 10.1006/dbio.2002.0645 PMID: 11977981.
61. Takamoto N, Zhao B, Tsai SY, DeMayo FJ. Identification of Indian hedgehog as a progesteroneresponsive gene in the murine uterus. Mol Endocrinol. 2002; 16(10):2338-48. doi: 10.1210/me.2001-0154 PMID: 12351698.
62. Hay-Koren A, Caspi M, Zilberberg A, Rosin-Arbesfeld R. The EDD E3 ubiquitin ligase ubiquitinates and up-regulates beta-catenin. Mol Biol Cell. 2011; 22(3):399-411. Epub 2010/12/02. doi: 10.1091/mbc. E10-05-0440 PMID: 21118991; PubMed Central PMCID: PMC3031469.
63. Wang G, Tang X, Chen Y, Cao J, Huang Q, Ling X, et al. Hyperplastic discs differentially regulates the transcriptional outputs of hedgehog signaling. Mech Dev. 2014; 133:117-25. doi: 10.1016/j.mod.2014. 05.002 PMID: 24854243; PubMed Central PMCID: PMCPMC4351657.
64. Sutherland C. What Are the bona fide GSK3 Substrates? Int J Alzheimers Dis. 2011; 2011:505607. Epub 2011/06/02. doi: 10.4061/2011/505607 PMID: 21629754; PubMed Central PMCID: PMC3100594.
65. Amano K, Ichida F, Sugita A, Hata K, Wada M, Takigawa Y, et al. MSX2 stimulates chondrocyte maturation by controlling Ihh expression. The Journal of biological chemistry. 2008; 283(43):29513-21.Epub 2008/08/07. doi: 10.1074/jbc.M803681200 PMID: 18682398; PubMed Central PMCID: PMC2662030.
66. Yoshida CA, Yamamoto H, Fujita T, Furuichi T, Ito K, Inoue K, et al. Runx2 and Runx3 are essential for chondrocyte maturation, and Runx2 regulates limb growth through induction of Indian hedgehog. Genes & development. 2004; 18(8):952-63. Epub 2004/04/27. doi: 10.1101/gad.1174704 PMID: 15107406; PubMed Central PMCID: PMC395853.
67. Wang W, Lian N, Li L, Moss HE, Perrien DS, Elefteriou F, et al. Atf4 regulates chondrocyte proliferation and differentiation during endochondral ossification by activating Ihh transcription. Development. 2009; 136(24):4143-53. Epub 2009/11/13. doi: 10.1242/dev.043281 PMID: 19906842; PubMed Central PMCID: PMC2781050.
68. Charite J, McFadden DG, Olson EN. The bHLH transcription factor dHAND controls Sonic hedgehog expression and establishment of the zone of polarizing activity during limb development. Development. 2000; 127(11):2461-70. PMID: 10804186.
69. Zhang Z, Sui P, Dong A, Hassell J, Cserjesi P, Chen YT, et al. Preaxial polydactyly: interactions among ETV, TWIST1 and HAND2 control anterior-posterior patterning of the limb. Development. 2010; 137 (20):3417-26. doi: 10.1242/dev.051789 PMID: 20826535; PubMed Central PMCID: PMCPMC2947755.
70. Lettice LA, Williamson I, Wiltshire JH, Peluso S, Devenney PS, Hill AE, et al. Opposing functions of the ETS factor family define Shh spatial expression in limb buds and underlie polydactyly. Dev Cell. 2012; 22(2):459-67. doi: 10.1016/j.devcel.2011.12.010 PMID: 22340503; PubMed Central PMCID: PMCPMC3314984.
71. Price MA, Kalderon D. Proteolysis of the Hedgehog signaling effector Cubitus interruptus requires phosphorylation by Glycogen Synthase Kinase 3 and Casein Kinase 1. Cell. 2002; 108(6):823-35.Epub 2002/04/17. PMID: 11955435.
72. Jia J, Amanai K, Wang G, Tang J, Wang B, Jiang J. Shaggy/GSK3 antagonizes Hedgehog signalling by regulating Cubitus interruptus. Nature. 2002; 416(6880):548-52. Epub 2002/03/26. doi: 10.1038/ nature733 PMID: 11912487.
73. Byrd N, Becker S, Maye P, Narasimhaiah R, St-Jacques B, Zhang X, et al. Hedgehog is required for murine yolk sac angiogenesis. Development. 2002; 129(2):361-72. PMID: 11807029.
74. Frisdal A, Trainor PA. Development and evolution of the pharyngeal apparatus. Wiley Interdiscip Rev Dev Biol. 2014; 3(6):403-18. doi: 10.1002/wdev.147 PMID: 25176500; PubMed Central PMCID: PMCPMC4199908.
75. Lettice LA, Williamson I, Devenney PS, Kilanowski F, Dorin J, Hill RE. Development of five digits is controlled by a bipartite long-range cis-regulator. Development. 2014; 141(8):1715-25. doi: 10.1242/dev.095430 PMID: 24715461; PubMed Central PMCID: PMCPMC3978833.
76. Gao B, Hu J, Stricker S, Cheung M, Ma G, Law KF, et al. A mutation in Ihh that causes digit abnormalities alters its signalling capacity and range. Nature. 2009; 458(7242):1196-200. doi: 10.1038/ nature07862 PMID: 19252479.
77. Zhang XM, Ramalho-Santos M, McMahon AP. Smoothened mutants reveal redundant roles for Shh and Ihh signaling including regulation of L/R asymmetry by the mouse node. Cell. 2001; 105(6):781-92. PMID: 11440720.