The secreted factor Sonic hedgehog (SHH) is both required for and sufficient to induce multiple developmental processes, including ventralization of the CNS, branching morphogenesis of the lungs and anteroposterior patterning of the limbs. Based on analogy to the Drosophila Hh pathway, the multiple GLI transcription factors in vertebrates are likely to both transduce SHH signaling and repress Shh transcription. In order to discriminate between overlapping versus unique requirements for the three Gli genes in mice, we have produced a Gli1 mutant and analyzed the phenotypes of Gli1/Gli2 and Gli1/3 double mutants. Gli3(xt) mutants have polydactyly and dorsal CNS defects associated with ectopic Shh expression, indicating GLI3 plays a role in repressing Shh. In contrast, Gli2 mutants have five digits, but lack a floorplate, indicating that it is required to transduce SHH signaling in some tissues. Remarkably, mice homozygous for a Gli1(zfd)mutation that deletes the exons encoding the DNA-binding domain are viable and appear normal. Transgenic mice expressing a GLI1 protein lacking the zinc fingers can not induce SHH targets in the dorsal brain, indicating that the Gli1(zfd)allele contains a hypomorphic or null mutation. Interestingly, Gli1(zfd/zfd);Gli2(zfd/+), but not Gli1(zfd/zfd);Gli3(zfd/+) double mutants have a severe phenotype; most Gli1(zfd/zfd);Gli2(zfd/+) mice die soon after birth and all have multiple defects including a variable loss of ventral spinal cord cells and smaller lungs that are similar to, but less extreme than, Gli2(zfd/zfd) mutants. Gli1/Gli2 double homozygous mutants have more extreme CNS and lung defects than Gli1(zfd/zfd);Gli2(zfd/+) mutants, however, in contrast to Shh mutants, ventrolateral neurons develop in the CNS and the limbs have 5 digits with an extra postaxial nubbin. These studies demonstrate that the zinc-finger DNA-binding domain of GLI1 protein is not required for SHH signaling in mouse. Furthermore, Gli1 and Gli2, but not Gli1 and Gli3, have extensive overlapping functions that are likely downstream of SHH signaling.
REFERENCES
Alexandre
C.
, Jacinto
A.
, Ingham
P. W.
(1996
) Transcriptional activation of hedgehog target genes in Drosophila is mediated directly by the Cubitus interruptus protein, a member of the GLI family of zinc finger DNA-binding proteins.
Genes Dev
10
, 2003
–2013
Aza-Blanc
P.
, Ramírez-Weber
F.-A.
, Laget
M.-P.
, Schwartz
C.
, Kornberg
T. B.
(1997
) Proteolysis that is inhibited by hedgehog targets cubitus interruptus protein to the nucleus and converts it to a repressor.
Cell
89
, 1043
–1053
Basler
K.
, Struhl
G.
(1994
) Compartment boundaries and the control of Drosophila limb pattern by hedgehog protein.
Nature
368
, 208
–214
Bellusci
S.
, Furuta
Y.
, Rush
M. G.
, Henderson
R.
, Winnier
G.
, Hogan
B. L.
(1997
) Involvement of Sonic hedgehog (Shh) in mouse embryonic lung growth and morphogenesis.
Development
124
, 53
–63
Bitgood
M.
, Shen
L.
, McMahon
A.
(1996
) Sertoli cell signaling by Desert hedgehog regulates the male germline.
Current Biol
6
, 298
–304
Bitgood
M. J.
, McMahon
A. P.
(1995
) Hedgehog an Bmp Genes Are Coexpressed at Many Diverse Sites of Cell-Cell Interaction in the Mouse Embryo.
Dev. Biol
172
, 126
–138
Borycki
A.
, Brunk
B.
, Tajbakhsh
S.
, Buckingham
M.
, Chiang
C.
, Emerson
C. P.
(1999
) Sonic hedgehog controls epaxial muscle determination through Myf5 activation.
Development
126
, 4053
–4063
Buscher
D.
, Bosse
B.
, Heymer
J.
, Ruther
U.
(1997
) Evidence for genetic control of Sonic hedgehog by Gli 3 in mouse limb development.
Mech. Dev
62
, 175
–182
Chiang
C.
, Litingtung
Y.
, Lee
E.
, Young
K. E.
, Corden
J. L.
, Westphal
H.
, Beachy
P. A.
(1996
) Cyclopia and defective axial patterning in mice lacking Sonic hedgehog gene function.
Nature
383
, 407
–413
Chiang
C.
, Swan
R. Z.
, Grachtchouk
M.
, Bolinger
M.
, Litingtung
Y.
, Robertson
E. K.
, Cooper
M. K.
, Gaffield
W.
, Westphal
H.
, Beachy
P. A.
(1999
) Essential role for Sonic hedgehog during hair follicle morphogenesis.
Dev. Biol
205
, 1
–9
Dai
P.
, Akimaru
H.
, Tanaka
Y.
, Maekawa
T.
, Nakafuku
M.
, Ishii
S.
(1999
) Sonic Hedgehog-induced activation of the Gli1 promoter is mediated by GLI3.
J. Biol. Chem
274
, 8143
–8152
Ding
Q.
, Motoyama
J.
, Gasca
S.
, Mo
R.
, Sasaki
H.
, Rossant
J.
, Hui
C. C.
(1998
) Diminished Sonic hedgehog signaling and lack of floor plate differentiation in Gli2 mutant mice.
Development
125
, 2533
–2543
Dominguez
M.
, Brunner
M.
, Hafen
E.
, Basler
K.
(1996
) Sending and Receiving the Hedgehog Signal: Control by the Drosophila Gli Protein Cubitus interruptus.
Science
272
, 1621
–1625
Echelard
Y.
, Epstein
D. J.
, St-Jacques
B.
, Shen
L.
, Mohler
J.
, McMahon
J. A.
, McMahon
A. P.
(1993
) Sonic hedgehog, a member of a family of putative signaling molecules, is implicated in the regulation of CNS polarity.
Cell
75
, 1417
–1430
Epstein
D. J.
, Marti
E.
, Scott
M. P.
, McMahon
A. P.
(1996
) Antagonizing cAMP-dependent protein kinase A in the dorsal CNS activates a conserved Sonic hedgehog signaling pathway.
Development
122
, 2885
–2894
Epstein
D. J.
, McMahon
A. P.
, Joyner
A. L.
(1999
) Regionalization of Sonic hedgehog transcription along the anteroposterior axis of the mouse central nervous system is regulated by Hnf3-dependent and-independent mechanisms.
Development
126
, 281
–292
Ericson
J.
, Thor
S.
, Edlund
T.
, Jessell
T. M.
, Yamada
T.
(1992
) Early stages of motor neuron differentiation revealed by expression of homeobox gene Islet-1.
Science
256
, 1555
–1560
Franz
T.
(1994
) Extra-Toes (Xt) homozygous mutant mice demonstrate a role for the Gli-3 gene in the development of the forebrain.
Acta Anat
150
, 38
–44
Goodrich
L. V.
, Johnson
R. L.
, Milenkovic
L.
, McMahon
J. A.
, Scott
M. P.
(1996
) Conservation of the hedgehog/patched signaling pathway from flies to mice: induction of a mouse patched gene by Hedgehog.
Genes Dev
10
, 301
–312
Gossler
A.
, Balling
R.
(1992
) The molecular and genetic analysis of mouse development.
Eur. J. Biochem
204
, 5
–11
Graves
L. E.
, Segal
S.
, Goodwin
E. B.
(1999
) TRA-1 regulates the cellular distribution of the tra-2 mRNA in C. elegans.
Nature
399
, 802
–805
Grindley
J. C.
, Bellusci
S.
, Perkins
D.
, Hogan
B. L.
(1997
) Evidence for the involvement of the Gli gene family in embryonic mouse lung development.
Dev. Biol
188
, 337
–348
Grove
E. A.
, Tole
S.
, Limon
J.
, Yip
L.
, Ragsdale
C. W.
(1998
) Thehem of the embryonic cerebral cortex is defined by the expression of multiple Wnt genes and is compromised in Gli3-deficient mice.
Development
125
, 2315
–2325
Hepker
J.
, Wang
Q. T.
, Motzny
C. K.
, Holmgren
R.
, Orenic
T. V.
(1997
) Drosophila cubitus interruptus forms a negative feedback loop with patched and regulates expression of Hedgehog target genes.
Development
124
, 549
–558
Hui
C. C.
, Joyner
A. L.
(1993
) A mouse model of Greig cephalopolysyndactyly syndrome: the extra-toes mutation contains an intragenic deletion of the Gli3 gene.
Nature Genet
3
, 241
–246
Hui
C. C.
, Slusarski
D.
, Platt
K.
, Homgren
R.
, Joyner
A.
(1994
) Expression of three mouse homologs of the Drosophila segment polarity gene cubitus interrupus, Gli, Gli-2, and Gli-3, in ectoderm and mesoderm-derived tissues suggests multiple roles during postimplantation development.
Dev. Biol
162
, 402
–413
Hynes
M.
, Porter
J.
(1995
) Induction of midbrain dopaminergic neurons by Sonic Hedgehog.
Neuron
15
, 35
–44
Hynes
M.
, Stone
D. M.
, Dowd
M.
, Pitts-Meek
S.
, Goddard
A.
, Gurney
A.
, Rosenthal
A.
(1997
) Control of cell pattern in the neural tube by the zinc finger transcription factor and oncogene Gli-1.
Neuron
19
, 15
–26
Ingham
P.
, Taylor
A.
, Nakano
Y.
(1991
) Role of the Drosophila patched gene in positional signalling.
Nature
353
, 184
–187
Johnson
R.
, Grenier
J.
, Scott
M.
(1995
) patched overexpression alters wing disc size and pattern: transcriptional and post-transcriptional effects on hedgehog targets.
Development
121
, 4161
–4170
Joyner
A.
, Kornberg
T.
, Coleman
K. G.
, Cox
D.
, Martin
G. R.
(1985
) Expression during embryogenesis of a mouse gene with sequence homology to the Drosophila engrailed gene.
Cell
43
, 29
–37
Kinzler
K. W.
, Ruppert
J. M.
, Bigner
S. H.
, Vogelstein
B.
(1988
) The GLI gene is a member of the Kruppel family of zinc finger proteins.
Nature
332
, 371
–374
Kinzler
K. W.
, Vogelstein
B.
(1990
) The GLI gene encodes a nuclear protein which binds specific sequences in the human genome.
Mol. Cell Biol
10
, 634
–642
Knecht
A. K.
, Harland
R. M.
(1997
) Mechanisms of dorsal-ventral patterning in noggin-induced neural tissue.
Development
124
, 2477
–2488
Krishnan
V.
, Pereira
F. A.
, Qui
Y.
, Chen
C.-H.
, Beachy
P. A.
, Tsai
S. Y.
, Tsai
M.-J.
(1997
) Mediation of Sonic Hedgehog-induced expression of COUP-TFII by a protein phosphatase.
Science
278
, 1947
–1950
Lee
J.
, Platt
K. A.
, Censullo
P.
, Ruiz i Altaba
A.
(1997
) Gli1 is a target of Sonic hedgehog that induces ventral neural tube development.
Development
124
, 2537
–2552
Lessing
D.
, Nusse
R.
(1998
) Expression of wingless in the Drosophila embryo: a conserved cis-acting element lacking conserved Ci-binding sites is required for patched-mediated repression.
Development
125
, 1469
–1476
Lewis
K. E.
, Drossopoulou
G.
, Paton
I. R.
, Morrice
D. R.
, Robertson
K. E.
, Burt
D. W.
, Ingham
P. W.
, Tickle
C.
(1999
) Expression of ptc and gli genes in talpid3 suggests bifurcation in Shh pathway.
Development
126
, 2397
–2407
Litingtung
Y.
, Lei
L.
, Westphal
H.
, Chiang
C.
(1998
) Sonic hedgehog is essential to foregut development.
Nat. Genet
20
, 58
–61
Liu
C. Z.
, Yang
J. T.
, Yoon
J. W.
, Villavicencio
E.
, Pfendler
K.
, Walterhouse
D.
, Iannaccone
P.
(1998
) Characterization of the promoter region and genomic organization of GLI, a member of the Sonic hedgehog-Patched signaling pathway.
Gene
209
, 1
–11
Marigo
V.
, Johnson
R.
, Vortkamp
A.
, Tabin
C.
(1996
) Sonic hedgehog differentially regulates expression of GLI and GLI3 during limb development.
Dev. Biol
180
, 273
–283
Marine
J. C.
, Bellefroid
E. J.
, Pendeville
H.
, Martial
J. A.
, Pieler
T.
(1997
) A role for Xenopus Gli-type zinc finger proteins in the early embryonic patterning of mesoderm and neuroectoderm.
Mech. Dev
63
, 211
–225
Matise
M. P.
, Epstein
D. J.
, Park
H. L.
, Platt
K. A.
, Joyner
A. L.
(1998
) Gli2 is required for induction of floor plate and adjacent cells, butnot most ventral neurons in the mouse central nervous system.
Development
125
, 2759
–2770
Matise
M. P.
, Lustig
M.
, Sakurai
T.
, Grumet
M.
, Joyner
A. L.
(1999
) Ventral midline cells are required for the local control of commissural axon gudance in the mouse spinal cord.
Development
126
, 3649
–3659
Melton
D. W.
(1994
) Gene targeting in the mouse.
BioEssays
16
, 633
–638
Mering
C. v.
, Basler
K.
(1999
) Distinct and regulated activities of human Gli proteins in Drosophila.
Current Biol
9
, 1319
–1322
Methot
N.
, Basler
K.
(1999
) Hedgehog controls limb development by regulating the activities of distinct transcriptional activator and repressor forms of Cubitus interruptus.
Cell
96
, 819
–831
Mo
R.
, Freer
A. M.
, Zinyk
D. L.
, Crackower
M. A.
, Michaud
J.
, Heng
H. H.
, Chik
K. W.
, Shi
X. M.
, Tsui
L. C.
, Cheng
S. H.
(1997
) Specific and redundant functions of Gli2 and Gli3 zinc finger genes in skeletal patterning and development.
Development
124
, 113
–123
Mohler
J.
(1988
) Requirements for hedgehog, a segmental polarity gene, in patterning larval and adult cuticle of Drosophila.
Genetics
120
, 1061
–1072
Monnier
V.
, Dussillol
F.
, Alves
G.
, Lamour-Isnard
C.
, Plessis
A.
(1998
) Suppressor of fused links fused and Cubitus interruptus on the hedgehog signalling pathway.
Current Biol
8
, 583
–586
Motoyama
J.
, Liu
J.
, Mo
R.
, Ding
Q.
, Post
M.
, Hui
C. C.
(1998
) Essential function of Gli2 and Gli3 in the formation of lung, trachea and oesophagus [see comments].
Nat. Genet
20
, 54
–57
Nagy
A.
, Rossant
J.
, Nagy
R.
, Abramow
N. W.
, Roder
J. C.
(1993
) Derivation of completely cell culture-derived mice from early-passage embryonic stem cells.
Proc. Natl Acad. Sci. USA
90
, 8424
–8428
Ohlmeyer
J. T.
, Kalderon
D.
(1998
) Hedgehog stimulates maturation of Cubitus interruptus into a labile transcriptional activator.
Nature
396
, 749
–753
Orenic
T. V.
, Slusarski
D. C.
, Kroll
K. L.
, Holmgren
R. A.
(1990
) Cloning and characterization of the segment polarity gene cubitus interruptus dominant of Drosophila.
Genes Dev
4
, 1053
–1067
Parr
B. A.
, Shea
M. J.
, Vassileva
G.
, McMahon
A. P.
(1993
) Mouse Wnt genes exhibit discrete domains of expression in the early embryonic CNS and limb buds.
Development
119
, 247
–261
Pavletich
N. P.
, Pabo
C. O.
(1993
) Crystal structure of a five-finger GLI-DNA complex: new perspectives on zinc fingers.
Science
261
, 1701
–1707
Pepicelli
C. V.
, Lewis
P. M.
, McMahon
A. P.
(1998
) Sonic hedgehog regulates branching morphogenesis in the mammalian lung.
Curr. Biol
8
, 1083
–1086
Platt
K. A.
, Michaud
J.
, Joyner
A. L.
(1997
) Expression of the mouse Gli and Ptc genes is adjacent to embryonic sources of hedgehog signals suggesting a conservation of pathways between flies and mice.
Mech. Dev
62
, 121
–135
Riddle
R. D.
, Johnson
R. L.
, Laufer
E.
, Tabin
C.
(1993
) Sonic hedgehog mediates the polarizing activity of the ZPA.
Cell
75
, 1401
–1416
Robbins
D. J.
, Nybakken
K. E.
, Kobayashi
R.
, Sisson
J. C.
, Bishop
J. M.
, Therond
P. P.
(1997
) Hedgehog elicits signal transduction by means of a large complex containing the kinesin-related protein Costal2.
Cell
90
, 225
–234
Roths
J. B.
, Foxworth
W. B.
, McArthur
M. J.
, Montgomery
C. A.
, Kier
A. B.
(1999
) Spontaneous and engineered mutant mice as models for experimental and comparative pathology: history, comparison, and developmental technology.
Laboratory Animal Science
49
, 12
–34
Ruiz i Altaba
A.
(1998
) Combinatorial Gli gene function in floor plate and neuronal induction by Sonic hedgehog.
Development
125
, 2203
–2212
Ruiz i Altaba
A.
(1999
) Gli proteins encode context-dependent positive and negative functions: implications for development and disease.
Development
126
, 3205
–3216
Ruiz i Altaba
A.
, Jessell
T.
, Roelink
H.
(1995
) Restrictions to floor plate induction by hedgehog and winged-helix genes in the neural tube of frog embryos.
Molec. Cell. Neurosci
6
, 106
–121
Ruiz i Altaba
A.
, Placzek
M.
, Baldassare
M.
, Dodd
J.
, Jessell
T. M.
(1995
) Early stages of notchord and floor plate development in the chick embryo defined by normal and induced expression of HNF-3.
Dev. Biol
170
, 299
–313
Sasaki
H.
, Hui
C.-c.
, Nakafuku
M.
, Kondoh
H.
(1997
) A binding site for Gli proteins is essential for HNF-3 floor plate enhancer activity in transgenics and can respond to Shh in vitro.
Development
124
, 1313
–1322
Sasaki
H.
, Nishizaki
Y.
, Hui
C. C.
, Nakafuku
M.
, Kondoh
H.
(1999
) Regulation of Gli2 and Gli3 activities by and amino-terminal repression domain: implication of Gli2 and Gli3 as primary mediators of Shh signaling.
Development
126
, 3915
–3924
Schimmang
T.
, Lemaistre
M.
, Vortkamp
A.
, Ruther
U.
(1992
) Expression of the zinc finger gene Gli3 is affected in the morphogenetic mouse mutant extra-toes (Xt).
Development
116
, 799
–804
Schwartz
C.
, Locke
J.
, Nishida
C.
, Kornberg
T. B.
(1995
) Analysis of cubitus interruptus regulation in Drosophila embryos and imaginal disks.
Development
121
, 1625
–1635
Sisson
J. C.
, Ho
K. S.
, Suyama
K.
, Scott
M. P.
(1997
) Costal2, a novel kinesin-related protein in the Hedgehog signaling pathway.
Cell
90
, 235
–245
St-Jacques
B.
, Dassule
H. R.
, Karavanova
I.
, Botchkarev
V. A.
, Li
J.
, Danielian
P. S.
, McMahon
J. A.
, Lewis
P. M.
, Paus
R.
, McMahon
A. P.
(1998
) Sonic hedgehog signaling is essential for hair development.
Curr. Biol
8
, 1058
–1068
Tabata
T.
, Eaton
S.
, Kornberg
T. B.
(1992
) The Drosophila hedgehog gene is expressed specifically in posterior compartment cells and is a target of engrailed regulation.
Genes Dev
6
, 2635
–2645
Tabata
T.
, Kornberg
T. B.
(1994
) Hedgehog is a signaling protein with a key role in patterning Drosophila imaginal discs.
Cell
76
, 89
–102
Takebe
Y.
, Seiki
M.
, Fujisawa
J.-I.
, Hoy
P.
, Yokota
K.
, Arai
K.-I.
, Yoshida
M.
, Arai
N.
(1988
) SRpromoter: an efficient and versatile mammalian cDNA expression system composed of the Simian Virus 40 early promoter and the R-U5 segment of human T-cell leukemia virus type 1 long terminal repeat.
Molec. Cell. Biol
8
, 466
–472
Takuma
N.
, Sheng
H. Z.
, Furuta
Y.
, Ward
J. M.
, Sharma
K.
, Hogan
B.
, Pfaff
S. L.
, Westphal
H.
, Kimura
S.
, Mahon
K. A.
(1998
) Formation of Rathke's pouch requires dual induction from the diencephalon.
Development
125
, 4835
–4840
Toresson
H.
, Mata de Urquiza
A.
, Fagerstrom
C.
, Perlmann
T.
, Campbell
K.
(1999
) Retinoids are produced by glia in the lateral ganglionic eminence and regulate striatal neuron differentiation.
Development
126
, 1317
–1326
Walterhouse
D.
, Ahmed
M.
, Slusarski
D.
, Kalamaras
J.
, Boucher
D.
, Holmgren
R.
, Iannaccone
P.
(1993
) gli, a zinc finger transcription factor and oncogene, is expressed during normal mouse development.
Dev. Dyn
196
, 91
–102
Wood
S. A.
, Allen
N. D.
, Rossant
J.
, Auerbach
A.
, Nagy
A.
(1993
) Non-injection methods for the production of embryonic stem cell-embryo chimaeras.
Nature
365
, 87
–89
Zuniga
A.
, Haramis
A.-P. G.
, McMahon
A. P.
, Zeller
R.
(1999
) Signal relay by BMP antagonism controls the SHH/FGF4 feedback loop in vertebrate limb buds.
Nature
401
, 598
–602
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