{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T06:12:18Z","timestamp":1791353538628,"version":"4.3.1"},"reference-count":48,"publisher":"The Company of Biologists","issue":"13","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2005,7,1]]},"abstract":"<jats:p>Intraflagellar transport (IFT) is an active event in which cargo is transported along microtubules by motor proteins such as kinesin and dynein. IFT proteins are required for the formation and maintenance of flagella and cilia. We have previously shown that mouse mutants for two IFT proteins, IFT88 and IFT172, as well as Kif3a, a subunit of mouse kinesin 2, exhibit ventral spinal cord patterning defects that appear to result from reduced hedgehog(Hh) signaling. Although genetic epistasis experiments place IFT proteins downstream of the Hh receptor and upstream of the Gli transcription factors,the mechanism by which IFT regulates Gli function is unknown.<\/jats:p><jats:p>The developing limb provides an excellent system to study Hh signaling, in particular as it allows a biological and molecular readout of both Gli activator and repressor function. Here we report that homozygous mutants for flexo (Fxo), a hypomorphic allele of mouse IFT88 generated in our ENU mutagenesis screen, exhibit polydactyly in all four limbs. Molecular analysis indicates that expression domains of multiple posteriorly restricted genes are expanded anteriorly in the mutant limbs, similar to loss of Gli3 transcriptional repressor function. Sonic hedgehog (Shh) expression is normal, yet Ptch1 and Gli1, two known targets of Hh signaling, are greatly reduced, consistent with loss of Shh signaling. Expression of Gli3 and Hand2 in the mutant limb indicates that the limb prepattern is abnormal. In addition, we show that partial loss-of-function mutations in another mouse IFT gene, Ift52(Ngd5), result in similar phenotypes and abnormal Hh signaling as Fxo, indicating a general requirement for IFT proteins in Hh signaling and patterning of multiple organs. Analysis of Ift88 and Shh double mutants indicates that, in mouse, IFT proteins are required for both Gli activator and repressor functions, and Gli proteins are insensitive to Hh ligand in the absence of IFT proteins. Finally, our biochemical studies demonstrate that IFT proteins are required for proteolytic processing of Gli3 in mouse embryos. In summary, our results indicate that IFT function is crucial in the control of both the positive and negative transcriptional activities of Gli proteins, and essential for Hh ligand-induced signaling cascade.<\/jats:p>","DOI":"10.1242\/dev.01894","type":"journal-article","created":{"date-parts":[[2005,6,2]],"date-time":"2005-06-02T00:25:37Z","timestamp":1117671937000},"page":"3103-3111","source":"Crossref","is-referenced-by-count":494,"title":["Mouse intraflagellar transport proteins regulate both the activator and repressor functions of Gli transcription factors"],"prefix":"10.1242","volume":"132","author":[{"given":"Aimin","family":"Liu","sequence":"first","affiliation":[{"name":"Department of Pediatrics, University of Colorado at Denver and Health Sciences Center, Mailstop 8322, Box 6511, Aurora, CO 80045, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Baolin","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Genetic Medicine, Cornell University Weill College of Medicine,New York, NY 10021,USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lee A.","family":"Niswander","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, University of Colorado at Denver and Health Sciences Center, Mailstop 8322, Box 6511, Aurora, CO 80045, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"237","reference":[{"key":"2021042605095971200_REF1","doi-asserted-by":"crossref","unstructured":"Bai, C. B., Stephen, D. and Joyner, A. L.(2004). All mouse ventral spinal cord patterning by hedgehog is Gli dependent and involves an activator function of Gli3. Dev. Cell6,103-115.","DOI":"10.1016\/S1534-5807(03)00394-0"},{"key":"2021042605095971200_REF2","doi-asserted-by":"crossref","unstructured":"Bak, M., Hansen, C., Tommerup, N. and Larsen, L. A.(2003). The Hedgehog signaling pathway-implications for drug targets in cancer and neurodegenerative disorders. Pharmacogenomics4,411-429.","DOI":"10.1517\/phgs.4.4.411.22751"},{"key":"2021042605095971200_REF3","doi-asserted-by":"crossref","unstructured":"Baker, S. A., Freeman, K., Luby-Phelps, K., Pazour, G. J. and Besharse, J. C. (2003). IFT20 links kinesin II with a mammalian intraflagellar transport complex that is conserved in motile flagella and sensory cilia. J. Biol. Chem.278,34211-34218.","DOI":"10.1074\/jbc.M300156200"},{"key":"2021042605095971200_REF4","doi-asserted-by":"crossref","unstructured":"Buscher, D., Bosse, B., Heymer, J. and Ruther, U.(1997). Evidence for genetic control of Sonic hedgehogby Gli3 in mouse limb development. Mech. Dev.62,175-182.","DOI":"10.1016\/S0925-4773(97)00656-4"},{"key":"2021042605095971200_REF5","doi-asserted-by":"crossref","unstructured":"Chiang, C., Litingtung, Y., Lee, E., Young, K. E., Corden, J. L., Westphal, H. and Beachy, P. A. (1996). Cyclopia and defective axial patterning in mice lacking Sonic hedgehog gene function. Nature383,407-413.","DOI":"10.1038\/383407a0"},{"key":"2021042605095971200_REF6","doi-asserted-by":"crossref","unstructured":"Chiang, C., Litingtung, Y., Harris, M. P., Simandl, B. K., Li,Y., Beachy, P. A. and Fallon, J. F. (2001). Manifestation of the limb prepattern: limb development in the absence of sonic hedgehog function. Dev. Biol.236,421-435.","DOI":"10.1006\/dbio.2001.0346"},{"key":"2021042605095971200_REF7","doi-asserted-by":"crossref","unstructured":"Chuang, P. T. and McMahon, A. P. (1999). Vertebrate Hedgehog signalling modulated by induction of a Hedgehog-binding protein. Nature397,617-621.","DOI":"10.1038\/17611"},{"key":"2021042605095971200_REF8","doi-asserted-by":"crossref","unstructured":"Dai, P., Akimaru, H., Tanaka, Y., Maekawa, T., Nakafuku, M. and Ishii, S. (1999). Sonic Hedgehog-induced activation of the Gli1 promoter is mediated by GLI3. J. Biol. Chem.274,8143-8152.","DOI":"10.1074\/jbc.274.12.8143"},{"key":"2021042605095971200_REF9","doi-asserted-by":"crossref","unstructured":"Ding, Q., Motoyama, J., Gasca, S., Mo, R., Sasaki, H., Rossant,J. and Hui, C. C. (1998). Diminished Sonic hedgehog signaling and lack of floor plate differentiation in Gli2 mutant mice. Development125,2533-2543.","DOI":"10.1242\/dev.125.14.2533"},{"key":"2021042605095971200_REF10","doi-asserted-by":"crossref","unstructured":"Goodrich, L. V., Milenkovic, L., Higgins, K. M. and Scott, M. P. (1997). Altered neural cell fates and medulloblastoma in mouse patched mutants. Science277,1109-1113.","DOI":"10.1126\/science.277.5329.1109"},{"key":"2021042605095971200_REF11","doi-asserted-by":"crossref","unstructured":"Han, Y. G., Kwok, B. H. and Kernan, M. J.(2003). Intraflagellar transport is required in Drosophila to differentiate sensory cilia but not sperm. Curr. Biol.13,1679-1686.","DOI":"10.1016\/j.cub.2003.08.034"},{"key":"2021042605095971200_REF12","doi-asserted-by":"crossref","unstructured":"Huangfu, D., Liu, A., Rakeman, A. S., Murcia, N. S., Niswander,L. and Anderson, K. V. (2003). Hedgehog signalling in the mouse requires intraflagellar transport proteins. Nature426,83-87.","DOI":"10.1038\/nature02061"},{"key":"2021042605095971200_REF13","doi-asserted-by":"crossref","unstructured":"Hui, C. C. and Joyner, A. L. (1993). A mouse model of greig cephalopolysyndactyly syndrome: the extra-toesJ mutation contains an intragenic deletion of the Gli3 gene. Nat. Genet.3,241-246.","DOI":"10.1038\/ng0393-241"},{"key":"2021042605095971200_REF14","doi-asserted-by":"crossref","unstructured":"Ingham, P. W. and McMahon, A. P. (2001). Hedgehog signaling in animal development: paradigms and principles. Genes Dev.15,3059-3087.","DOI":"10.1101\/gad.938601"},{"key":"2021042605095971200_REF15","doi-asserted-by":"crossref","unstructured":"Izraeli, S., Lowe, L. A., Bertness, V. L., Good, D. J., Dorward,D. W., Kirsch, I. R. and Kuehn, M. R. (1999). The SIL gene is required for mouse embryonic axial development and left-right specification. Nature399,691-694.","DOI":"10.1038\/21429"},{"key":"2021042605095971200_REF16","doi-asserted-by":"crossref","unstructured":"Jia, J., Tong, C. and Jiang, J. (2003). Smoothened transduces Hedgehog signal by physically interacting with Costal2\/Fused complex through its C-terminal tail. Genes Dev.17,2709-2720.","DOI":"10.1101\/gad.1136603"},{"key":"2021042605095971200_REF17","doi-asserted-by":"crossref","unstructured":"Knezevic, V., De Santo, R., Schughart, K., Huffstadt, U.,Chiang, C., Mahon, K. A. and Mackem, S. (1997). Hoxd-12 differentially affects preaxial and postaxial chondrogenic branches in the limb and regulates Sonic hedgehog in a positive feedback loop. Development124,4523-4536.","DOI":"10.1242\/dev.124.22.4523"},{"key":"2021042605095971200_REF18","doi-asserted-by":"crossref","unstructured":"Kraus, P., Fraidenraich, D. and Loomis, C. A.(2001). Some distal limb structures develop in mice lacking Sonic hedgehog signaling. Mech. 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Neurosci.3, 979-985.","DOI":"10.1038\/79916"},{"key":"2021042605095971200_REF22","doi-asserted-by":"crossref","unstructured":"Litingtung, Y., Dahn, R. D., Li, Y., Fallon, J. F. and Chiang,C. (2002). Shh and Gli3 are dispensable for limb skeleton formation but regulate digit number and identity. Nature418,979-983.","DOI":"10.1038\/nature01033"},{"key":"2021042605095971200_REF23","doi-asserted-by":"crossref","unstructured":"Liu, A., Joyner, A. L. and Turnbull, D. H.(1998). Alteration of limb and brain patterning in early mouse embryos by ultrasound-guided injection of Shh-expressing cells. Mech. Dev.75,107-115.","DOI":"10.1016\/S0925-4773(98)00090-2"},{"key":"2021042605095971200_REF24","doi-asserted-by":"crossref","unstructured":"Liu, A., Losos, K. and Joyner, A. L. (1999). FGF8 can activate Gbx2 and transform regions of the rostral mouse brain into a hindbrain fate. Development126,4827-4838.","DOI":"10.1242\/dev.126.21.4827"},{"key":"2021042605095971200_REF25","doi-asserted-by":"crossref","unstructured":"Lum, L. and Beachy, P. A. (2004). The Hedgehog response network: sensors, switches, and routers. Science304,1755-1759.","DOI":"10.1126\/science.1098020"},{"key":"2021042605095971200_REF26","doi-asserted-by":"crossref","unstructured":"Lum, L., Zhang, C., Oh, S., Mann, R. K., von Kessler, D. P.,Taipale, J., Weis-Garcia, F., Gong, R., Wang, B. and Beachy, P. A.(2003). Hedgehog signal transduction via Smoothened association with a cytoplasmic complex scaffolded by the atypical kinesin,Costal-2. Mol. Cell12,1261-1274.","DOI":"10.1016\/S1097-2765(03)00426-X"},{"key":"2021042605095971200_REF27","doi-asserted-by":"crossref","unstructured":"Matise, M. P., Epstein, D. J., Park, H. L., Platt, K. A. and Joyner, A. L. (1998). Gli2 is required for induction of floor plate and adjacent cells, but not most ventral neurons in the mouse central nervous system. Development125,2759-2770.","DOI":"10.1242\/dev.125.15.2759"},{"key":"2021042605095971200_REF28","doi-asserted-by":"crossref","unstructured":"McCarthy, R. A., Barth, J. L., Chintalapudi, M. R., Knaak, C. and Argraves, W. S. (2002). Megalin functions as an endocytic sonic hedgehog receptor. J. Biol. Chem.277,25660-25667.","DOI":"10.1074\/jbc.M201933200"},{"key":"2021042605095971200_REF29","unstructured":"Morgan, B. A., Izpisua-Belmonte, J. C., Duboule, D. and Tabin,C. J. (1992). Targeted misexpression of Hox-4.6 in the avian limb bud causes apparent homeotic transformations. Nature358,236-239."},{"key":"2021042605095971200_REF30","doi-asserted-by":"crossref","unstructured":"Moyer, J. H., Lee-Tischler, M. J., Kwon, H. Y., Schrick, J. J.,Avner, E. D., Sweeney, W. E., Godfrey, V. L., Cacheiro, N. L., Wilkinson, J. E. and Woychik, R. P. (1994). Candidate gene associated with a mutation causing recessive polycystic kidney disease in mice. Science264,1329-1333.","DOI":"10.1126\/science.8191288"},{"key":"2021042605095971200_REF31","doi-asserted-by":"crossref","unstructured":"Niswander, L. (2003). Pattern formation: old models out on a limb. Nat. Rev. Genet.4, 133-143.","DOI":"10.1038\/nrg1001"},{"key":"2021042605095971200_REF32","doi-asserted-by":"crossref","unstructured":"Niswander, L. and Martin, G. R. (1992). Fgf-4 expression during gastrulation, myogenesis, limb and tooth development in the mouse. Development114,755-768.","DOI":"10.1242\/dev.114.3.755"},{"key":"2021042605095971200_REF33","doi-asserted-by":"crossref","unstructured":"Ogden, S. K., Ascano, M., Jr, Stegman, M. A., Suber, L. M.,Hooper, J. E. and Robbins, D. J. (2003). Identification of a functional interaction between the transmembrane protein Smoothened and the kinesin-related protein Costal2. Curr. Biol.13,1998-2003.","DOI":"10.1016\/j.cub.2003.10.004"},{"key":"2021042605095971200_REF34","doi-asserted-by":"crossref","unstructured":"Riddle, R. D., Johnson, R. L., Laufer, E. and Tabin, C.(1993). Sonic hedgehog mediates the polarizing activity of the ZPA. Cell75,1401-1416.","DOI":"10.1016\/0092-8674(93)90626-2"},{"key":"2021042605095971200_REF35","doi-asserted-by":"crossref","unstructured":"Rosenbaum, J. L. and Witman, G. B. (2002). Intraflagellar transport. Nat. Rev. Mol. Cell Biol.3, 813-825.","DOI":"10.1038\/nrm952"},{"key":"2021042605095971200_REF36","doi-asserted-by":"crossref","unstructured":"Ruel, L., Rodriguez, R., Gallet, A., Lavenant-Staccini, L. and Therond, P. P. (2003). Stability and association of Smoothened, Costal2 and Fused with Cubitus interruptus are regulated by Hedgehog. Nat. Cell Biol.5, 907-913.","DOI":"10.1038\/ncb1052"},{"key":"2021042605095971200_REF37","doi-asserted-by":"crossref","unstructured":"Sarpal, R., Todi, S. V., Sivan-Loukianova, E., Shirolikar, S.,Subramanian, N., Raff, E. C., Erickson, J. W., Ray, K. and Eberl, D. F.(2003). Drosophila KAP interacts with the kinesin II motor subunit KLP64D to assemble chordotonal sensory cilia, but not sperm tails. Curr. Biol.13,1687-1696.","DOI":"10.1016\/j.cub.2003.09.025"},{"key":"2021042605095971200_REF38","doi-asserted-by":"crossref","unstructured":"Sasaki, H., Nishizaki, Y., Hui, C., Nakafuku, M. and Kondoh,H. (1999). Regulation of Gli2 and Gli3 activities by an amino-terminal repression domain: implication of Gli2 and Gli3 as primary mediators of Shh signaling. Development126,3915-3924.","DOI":"10.1242\/dev.126.17.3915"},{"key":"2021042605095971200_REF39","doi-asserted-by":"crossref","unstructured":"Sekimizu, K., Nishioka, N., Sasaki, H., Takeda, H., Karlstrom,R. O. and Kawakami, A. (2004). The zebrafish iguana locus encodes Dzip1, a novel zinc-finger protein required for proper regulation of Hedgehog signaling. Development131,2521-2532.","DOI":"10.1242\/dev.01059"},{"key":"2021042605095971200_REF40","doi-asserted-by":"crossref","unstructured":"te Welscher, P., Fernandez-Teran, M., Ros, M. A. and Zeller,R. (2002a). Mutual genetic antagonism involving GLI3 and dHAND prepatterns the vertebrate limb bud mesenchyme prior to SHH signaling. Genes Dev.16,421-426.","DOI":"10.1101\/gad.219202"},{"key":"2021042605095971200_REF41","doi-asserted-by":"crossref","unstructured":"te Welscher, P., Zuniga, A., Kuijper, S., Drenth, T., Goedemans,H. J., Meijlink, F. and Zeller, R. (2002b). Progression of vertebrate limb development through SHH-mediated counteraction of GLI3. Science298,827-830.","DOI":"10.1126\/science.1075620"},{"key":"2021042605095971200_REF42","doi-asserted-by":"crossref","unstructured":"Timmer, J., Johnson, J. and Niswander, L.(2001). The use of in ovo electroporation for the rapid analysis of neural-specific murine enhancers. Genesis29,123-132.","DOI":"10.1002\/gene.1015"},{"key":"2021042605095971200_REF43","doi-asserted-by":"crossref","unstructured":"Wang, B., Fallon, J. F. and Beachy, P. A.(2000). Hedgehog-regulated processing of Gli3 produces an anterior\/posterior repressor gradient in the developing vertebrate limb. Cell100,423-434.","DOI":"10.1016\/S0092-8674(00)80678-9"},{"key":"2021042605095971200_REF44","doi-asserted-by":"crossref","unstructured":"Wang, Q. T. and Holmgren, R. A. (2000). Nuclear import of cubitus interruptus is regulated by hedgehog via a mechanism distinct from Ci stabilization and Ci activation. Development127,3131-3139.","DOI":"10.1242\/dev.127.14.3131"},{"key":"2021042605095971200_REF45","doi-asserted-by":"crossref","unstructured":"Wick, M. J., Ann, D. K. and Loh, H. H. (1995). Molecular cloning of a novel protein regulated by opioid treatment of NG108-15 cells. Brain Res. Mol. Brain Res.32,171-175.","DOI":"10.1016\/0169-328X(95)00090-F"},{"key":"2021042605095971200_REF46","doi-asserted-by":"crossref","unstructured":"Wolff, C., Roy, S., Lewis, K. E., Schauerte, H., Joerg-Rauch,G., Kirn, A., Weiler, C., Geisler, R., Haffter, P. and Ingham, P. W.(2004). iguana encodes a novel zinc-finger protein with coiled-coil domains essential for Hedgehog signal transduction in the zebrafish embryo. Genes Dev.18,1565-1576.","DOI":"10.1101\/gad.296004"},{"key":"2021042605095971200_REF47","doi-asserted-by":"crossref","unstructured":"Yang, Y., Guillot, P., Boyd, Y., Lyon, M. F. and McMahon, A. P. (1998). Evidence that preaxial polydactyly in the Doublefoot mutant is due to ectopic Indian Hedgehog signaling. Development125,3123-3132.","DOI":"10.1242\/dev.125.16.3123"},{"key":"2021042605095971200_REF48","doi-asserted-by":"crossref","unstructured":"Zhang, Q., Murcia, N. S., Chittenden, L. R., Richards, W. G.,Michaud, E. J., Woychik, R. P. and Yoder, B. K. (2003). 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