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First published online December 12, 2006
doi: 10.1242/10.1242/dev.02703
Research Report |
1 Developmental Skin Biology Unit, NIAMS, NIH, Bethesda, MD 20892, USA.
2 Department of Biomolecular and Biotechnological Sciences, University of Milan,
Via Celoria 26. 20133, Milan, Italy.
3 Dulbecco Telethon Institute c/o Istituto Tecnologie Biomediche CNR, 20100
Milan, Italy.
4 Department of Dermatology, University of Rome `Tor Vergata', Viale Oxford 81.
00133 Rome, Italy.
5 Division of Monoclonal Antibodies, CDER/FDA, Bethesda, MD 20892, USA.
6 Department of Structural and Functional Biology, University of Naples, Via
Cinzia 26. 80126 Naples, Italy.
Author for correspondence (e-mail:
morassom{at}mail.nih.gov)
Accepted 18 October 2006
SUMMARY
Ectodermal dysplasias (EDs) are a group of human pathological conditions
characterized by anomalies in organs derived from epithelial-mesenchymal
interactions during development. Dlx3 and p63 act as part of the
transcriptional regulatory pathways relevant in ectoderm derivatives, and
autosomal mutations in either of these genes are associated with human EDs.
However, the functional relationship between both proteins is unknown. Here,
we demonstrate that Dlx3 is a downstream target of p63. Moreover, we
show that transcription of Dlx3 is abrogated by mutations in the
sterile 
-motif (SAM) domain of p63 that are associated with
ankyloblepharon-ectodermal dysplasia-clefting (AEC) dysplasias, but not by
mutations found in ectrodactylyectodermal dysplasia-cleft lip/palate (EEC),
Limb-mammary syndrome (LMS) and split hand-foot malformation (SHFM)
dysplasias. Our results unravel aspects of the transcriptional cascade of
events that contribute to ectoderm development and pathogenesis associated
with p63 mutations.
Key words: Dlx3, p63, Transcription, Ectodermal dysplasias, Mouse development
INTRODUCTION
During embryonic development and organ formation, a series of signals
between epithelial cells and the underlying mesenchymal cells are the basis
for the formation of a variety of appendages and/or organs
(Pispa and Thesleff, 2003
).
Anomalies in epithelialmesenchymal-derived organs are characteristics of human
pathological conditions defined as ectodermal dysplasias (EDs)
(Priolo and Lagana, 2001).
Mutations in DLX3 and p63, among other genes, have been
directly linked with EDs. The Dlx and p63 families of transcriptional
effectors are essential for the development of the epidermis and/or embryonic
appendages (Panganiban and Rubenstein,
2002; Merlo et al.,
2003; Morasso and Radoja,
2005; Koster and Roop,
2004). Dlx3 expression has been detected in the hair
follicle, tooth, limb bud, branchial arches, labyrinthe layer of the placenta,
osteoblasts and epidermis (Morasso et al.,
1995; Morasso et al.,
1999; Hassan et al.,
2004). Here, we present evidence that Dlx3 is regulated
by p63 as part of a transcriptional regulatory pathway relevant to specific
EDs.
p63 regulates multiple signaling pathways, such as the bone morphogenetic
protein (BMP) and fibroblast growth factor (FGF) pathways
(Laurikkala et al., 2006;
Barbieri and Pietenpol, 2006).
Transgenic and knockout (KO) mouse models indicate that p63 has essential
roles in the development and maintenance of the stratified epidermis
(Yang et al., 1999;
Mills et al., 1999;
Koster et al., 2004;
Koster and Roop, 2004). The
p63 gene is transcribed from two distinct promoters, giving rise to
proteins that either contain (TAp63) or lack (
Np63) the amino terminal
transactivating domain. The TA and N isoforms both possess the
DNA-binding and oligomerization domains, and, by alternative splicing at the
3' end, produce isoforms with different C-termini, termed alpha
(), beta (ß) and gamma (
)
(Yang et al., 1998). The
isoforms contain a sterile motif (SAM) - a domain with reputed
importance in protein-protein interactions
(Qiao and Bowie, 2005). p63
isoforms act as transcriptional activators and/or repressors
(Ghioni et al., 2002;
King et al., 2003;
Wu et al., 2005), and bind to
two or more tandem repeats of RRRCWWGYYY, but preferentially activate the
RRRCGTGYYY sequence (Osada et al.,
2005).
Mutations in the p63 gene have been associated with EDs that
include ectrodactyly-ectodermal dysplasia-cleft lip/palate (EEC), limb-mammary
syndrome (LMS), split hand-foot malformation (SHFM) and
ankyloblepharon-ectodermal dysplasia-clefting (AEC) syndrome. There is a
correlation between the position of the mutation and the observed abnormal
phenotype (van Bokhoven et al.,
2001; McGrath et al.,
2001; van Bokhoven and
Brunner, 2002). Mutations in the DLX3 gene are linked to
tricho-dento-osseus (TDO) syndrome, which, like AEC, is characterized by
defects in the development of hair, teeth and bone, and by absence of overt
limb malformations (Price et al.,
1998).
Partial-overlapping mRNA expression and phenotypes of specific human malformations caused by molecular lesions in either p63 or DLX3 suggest that these genes are components of common signaling pathways during embryonic development. Here, we show that p63 is able to bind and transactivate Dlx3 both in vitro and in vivo. Mutant p63 proteins derived from AEC patients exhibit an impaired ability to transactivate Dlx3, indicating that the misregulation of the DLX3 gene is involved in the pathogenesis of human syndromes associated to AEC.
MATERIALS AND METHODS
DNA constructs
The -117 to +60 DNA fragment of the Dlx3 promoter
(Park and Morasso, 1999) was
inserted into the pGL3-Basic and pCAT-Basic vectors (Promega). Mutations in
the p63-binding sites of the Dlx3 promoter were obtained using the
ExSite Mutagenesis kit (Stratagene). The coding sequences for
Np63, Np63ß, and
Np63 were cloned into pBK-CMV (Stratagene). The
TAp63, TAp63ß and TAp63
constructs were a gift from E. Candi (University of Rome `Tor Vergata', Rome,
Italy) and G. Melino (University of Rome `Tor Vergata', Rome, Italy). The p63
mutants L518F, L518V and Q540L (AEC); E639X (SHFM);
FS525 (EEC); and G76W and AA (LMS) were
kindly provided by H. van Bokhoven (Radboud University, Nijmegan, The
Netherlands).
|
).
Normal human epidermal keratinocytes [NHEK; a gift of M. Simon (SUNY, Stony
Brook, NY, USA)] were derived from newborn foreskin and cultured in K-SFM
(Invitrogen). The human osteosarcoma Saos-2 cell line was maintained in RPMI
1640 and 10% FCS. The immortalized human keratinocyte HaCaT and H1299
non-small-lungcarcinoma cell lines were grown in DMEM and 10% FBS.
Transient transfection
Transient transfections of keratinocytes were performed with FuGENE6
(Roche) in a 1:3 ratio. PRL-SV40 vector was used as an internal control.
Luciferase activity was measured 24-36 hours after transfection using the
Dual-Luciferase Reporter Assay System (Promega). Transient transfections of
Saos-2 and H1299 cells were performed according to Calabrò et al.
(Calabrò et al., 2002).
CAT reaction was performed 48 hours after transfection using 90 µg of cell
extract. ß-Gal was used to normalize for transfection efficiency.
EMSA analysis
Nuclear extracts and EMSA analysis were carried out according to Park and
Morasso (Park and Morasso,
1999) using the Dlx3 p63site1+2 and non-specific competitor AP-2
binding site.
Chromatin immunoprecipitation assay
Chromatin immunoprecipitation (ChIP) was performed with chromatin from
mouse keratinocytes and H1299 cells transfected with TAp63 according to
Caretti et al. (Caretti et al.,
2004) using no antibody, IgG or p63 antibody (Santa Cruz, H137 and
4A4). Real-time PCR was performed using the Mx3000P System (Stratagene) and
SyberGreen MasterMix (Applied Biosystems) with independent DNA samples and the
following oligonucleotides: for mouse, Dlx3(F)
5'-GAGAAAGCGCGAGCGTGTTTTGCC-3' and Dlx3(R)
5'-CCGGCTGTCGGTCAGTCGCTGCGT-3'; for human, DLX3(F)
5'-AGAGAGGCGGAAGAGACGAG-3' and DLX3(R)
5'-GAGGAGGGAGGAGAGAAGGA-3'; and for JAG2(F)
5'-CAAGTGGTGAACAAGGGAGACT-3' and JAG2(R)
5'-ACTGCTGCCTTCTGGAAACTC-3'. Data are presented as fold
differences relative to input and values are obtained by IgG with the formula
2[(CtIgG-CtInput)-(CtAb-CtInput)], where Ct is threshold cycles,
IgG is normal rabbit IgG, Ab is specific antibody and Input is input genomic
DNA. ACHR amplification was performed as a control for H1299 transfected with
TAp63 using ACHR(F) 5'-TGCCTCGGGTGAACTAAGATG-3' and ACHR(R)
5'-GCCTCATTCGTCTTGGGAACT-3'.
Real-time PCR
For analysis of the expression of p63 in mouse keratinocytes and
embryonic tissues, the following oligonucleotides were used: TAp63(F)
5'-AGACAAGCGAGTTCCTCAGC-3', TAp63(R)
5'-TGCGGATACAATCCATGCTA-3', Np63(F)
5'-ATGTTGTACCTGGAAAACAATG-3', Np63(R)
5'-GATGGAGAGAGGGCATCAAA-3'. These oligonucleotides are designed in
regions of the mRNA common to all isoforms (, ß and ), and
do not distinguish between these variants.
For Dlx3 expression in mouse keratinocytes, the following oligonucleotides were used: Dlx3(F) 5'-ATTACAGCGCTCCTCAGCAT-3' and Dlx3(R) 5'-GCCTATAGGATCCCCCGTAG-3'. In embryonic tissues: Dlx3(F) 5'-CGTTTCCAGAAAGCCCGTA-3' and Dlx3(R) 5'-CGTGGAATGGGAAGATGTGT-3'. For normalization: GAPDH(F) 5'-TGTCAGCAATGCATCCTGCA-3' and GADPH(R) 5'-TGTATGCAGGGATGATGTTC-3'.
|
Immunohistochemistry and whole-mount in situ hybridization
Immunohistochemistry was performed on 11 µm cryostatic sections of E10.5
embryonic forelimbs. Sections were blocked with 10% goat serum in PBS for 40
minutes at room temperature. Antibodies used were: mouse monoclonal anti-p63
(4A4, 1:100, Santa Cruz) and rabbit anti-distal-less [pan-anti-Dlx, 1:100;
kindly provided by G. Boekhoff-Falk (University of Wisconsin Medical School,
WI, USA)]. As secondary antibody, anti-mouse-Cy2, anti-rabbit-Cy3 (1:100;
Jackson Immuno-Research) and Envision antirabbit HRP (Dako) were used.
Fluorescence micrographs were taken by confocal microscopy.
Whole-mount in situ hybridization was performed according to Acampora et
al. (Acampora et al., 1999) on
E10.5 p63 KO embryos [Brdm2 line of p63 KO kindly provided
by D. Roop (Baylor College of Medicine, Houston, USA)] using a Dlx3
probe (Morasso et al.,
1995).
RT-PCR
Total RNA from human Saos-2 and HaCaT cells was prepared with TRIZOL
(Roche). For reverse transcription (RT)-PCR, 3-4 µg of total RNA were
reverse-transcribed using SuperScript II (Invitrogen). The following
oligonucleotides were used: Dlx3(F) 5'-ACCTACGGAGCCTCCTACCG-3',
Dlx3(R) 5'-ACTCAGGTTCTGTGCGTGAT-3', p63(F)
5'-GTCTCCATCTTCATATGGTAAC-3', p63(R)
5'-CACACTGACTGTAGAGGCA-3', p63ß(F)
5'-GTCTCCATCTTCATATGGTAAC-3', p63ß(R)
5'-CTTGCCAAATCCTGACAATGCTGC-3', p63(F)
5'-GAGGATAGCATCAGAAAACAGCAAG-3' and p63(R)
5'-CTCCACAAGCTCATTCCTGAAGC-3'. For normalization: Cyclophillin(F)
5'-ATCACCATTGCTGACTGTGG-3', Cyclophillin(R)
5'-ACTCTGCAATCCAGCTAGGC-3', GAPDH(F)
5'-GTCTCCATCTTCATATGGTAA-3' and GAPDH(R)
5'-CCACAGTCCATGCCATCACT-3' were used.
RESULTS AND DISCUSSION
The existence of malformations caused by either p63 or
DLX3 gene mutations that translate to partially overlapping
phenotypes suggests that these genes are transcriptional effectors in common
signaling cascades regulating epidermal development. The severity of the
phenotype in p63-null mice suggests that it is a crucial upstream regulator of
these signaling pathways (Mills et al.,
1999; Yang et al.,
1999). Detailed analysis of the Dlx3 proximal promoter
region revealed a sequence with two p63-like overlapping binding sites
immediately upstream of the CCAAT box, located from -89 to -80 bp (site 1) and
from -84 to -75 bp (site 2) of the transcriptional start site
(Park and Morasso, 1999).
Because the expression patterns of p63 and Dlx3 overlap
throughout embryonic development (Morasso
and Radoja, 2005), we proceeded to test the ability of different
p63 isoforms to transactivate the Dlx3 promoter. The
Dlx3-117/+60 construct, which contains the two overlapping sites,
was transiently transfected into primary mouse keratinocytes in either the
absence or presence of expression vectors encoding TAp63, TAp63ß,
TAp63, Np63, Np63ß or Np63. The
TA isoforms activated the Dlx3-117/+60 promoter at a magnitude of
twelve-, three- and seven-fold, respectively, compared to normal activation,
whereas the exogenous expression of the N isoforms resulted in a two-
to four-fold greater transactivation compared with wild type
(Fig. 1A). These effects are
specific for p63 isoforms, because p53 did not transactivate the Dlx3
promoter (data not shown).
|
Np63 isoforms (Fig.
1C,D). Our results indicate that TAp63 induction of
Dlx3 is mediated through either of the two p63 sites, whereas
TAp63 required an intact regulatory region. These results are not
cell-type specific, because a similar profile was obtained using Saos-2 cells
(Fig. 1E).
Since Dlx3 is induced in keratinocytes cultured in 0.12 mM
Ca2+ (Park and Morasso,
1999), we compared the endogenous expression of the p63
isoforms with Dlx3 after 12- and 24-hours of 0.12 mM Ca2+
treatment (Fig. 2A-C). The
real-time PCR results showed a correlation between the upregulation of
Dlx3 and TAp63 and the downregulation of
Np63 mRNAs associated by Ca2+-induced
differentiation, and are consistent with the recent report by King et al.
(King et al., 2006). The
specificity of the PCR products was corroborated by sequencing (data not
shown). TAp63 proteins are found in normal adult tissues
(Nylander et al., 2002) and
during mouse embryonic development (Koster
et al., 2004) (also our own data). Findings of Koster et al.
(Koster et al., 2004) support
a role for TAp63 as a molecular switch for the initiation of epithelial
stratification. Our findings support a working model in which, once
transactivated by TAp63, Dlx3 will in turn regulate
the expression of terminal differentiation markers
(Morasso et al., 1996).
In order to demonstrate direct binding to the p63 region in the
Dlx3 promoter, we performed EMSA with a fragment comprising -89 to
-75 bp (Dlx3 p63 sites 1 and 2) using nuclear extracts from primary
keratinocytes (NE). A shift was detected
(Fig. 2D, lane NE), and the
complexes were competed with a specific competitor
(Fig. 2D, lane SC), but not
with a nonspecific DNA competitor (Fig.
2D, lane NC). We next evaluated whether p63 bound this region of
the Dlx3 promoter in vivo. In mouse keratinocytes, ChIP experiments
were performed with a p63-specific antibody and analyzed by regular PCR
(Fig. 2E) and real-time PCR
(Fig. 2F). The data shows that
p63 specifically binds to the Dlx3 promoter in vivo (eightfold higher
than with IgG control). Moreover, ChIP experiments on TAp63-transfected
H1299 cells, which are devoid of p63, demonstrated the direct binding of
TAp63 to the Dlx3 promoter (Fig.
2G).
To further explore the relationship of Dlx3 and p63 in vivo, we analyzed
their colocalization by immunofluorescence on E10.5 embryonic forelimb
sections with anti-p63 and anti-distal-less antibodies. The latter reagent
recognizes Dlx3 in the limb ectoderm. p63 and Dlx3 immunoreactivity were found
to colocalize in the same nuclei (Fig.
3A). Comparison of the expression of Dlx3, TAp63 and
Np63 in the limbs at embryonic stages E10.5, E11.5 and E12.5
was performed by real-time PCR. Between E11.5 and E12.5, the relative
abundance of both TAp63 and Dlx3 mRNA increased from three-
to eight-fold relative to their expression at E10.5 in the anterior (AL) and
posterior limbs (PL) (Fig. 3B),
whereas expression of Np63 was only moderately increased.
These results for p63 in the limb ectoderm of embryos at E10.5-E12.5 are in
agreement with reported data (Koster et
al., 2004) and show for the first time that a good correlation is
observed between the expressions of Dlx3 and TAp63.
To provide further evidence that p63 is an upstream regulator of Dlx3, we
studied the effect of p63 ablation on Dlx3 mRNA and protein
expression in the Brdm2 p63 KO mice
(Mills et al., 1999)
(Fig. 3C-D). As assessed by
immunohistochemistry, the abundance of Dlx3 protein was significantly reduced
in p63-KO limb ectoderm (Fig.
3C). Analysis by whole-mount in situ hybridization with a
Dlx3 antisense probe demonstrated that the absence of p63 led to a
downregulation of Dlx3 (Fig.
3D).
We next studied the clinical relevance of p63-mediated regulation of
Dlx3 expression. We examined the transcriptional activity of the
Dlx3 promoter in the presence of p63 mutants causative of human AEC,
EEC, SHFM or LMS (Fig. 4A) in
the H1299 cell line. The p63 AEC mutants - L518V, L518F and Q540L -
are all point substitutions within the SAM domain, present only in
TAp63 and Np63. The
p63FS mutant contains a mutation found in EEC that
generates a frameshift at amino acid 525, which leads to a premature stop. The
E639X mutation, in exon 14, was isolated in a SHFM patient, whereas the 2-bp
deletion, in exon 14 (AA), was present in one family with LMS. The G76W
mutation, in exon 3, was isolated from a LMS patient and affects all p63
isoforms. Co-transfection experiments were performed with expression vectors
encoding TAp63 and Np63 mutants. As shown in
Fig. 4B and 4C, the AEC mutants
failed to yield a significant level of reporter-gene expression despite having
intact amino terminal and DNA-binding domains. These results point to a
crucial role of the C-terminus of p63 on the regulation of
Dlx3 transcription. The LMS-derived mutants G76W and AA; as
well as the FSEEC mutant, SHFM-derived E639X TA and N
proteins; showed a similar mode of regulation of Dlx3-promoter
activity compared with the corresponding wild-type proteins
(Fig. 4B,C). A similar profile,
albeit with a lower amount of induction, was obtained upon transfection in
HaCaT cells (data not shown). The level of expression of the mutant proteins
was corroborated by immunoblot analysis with anti-p63 antibody
(Fig. 4D).
|
; Morasso and Radoja,
2005).
To determine the modulation of endogenous Dlx3 by p63, we used
Saos-2 cells to express wild-type and mutant p63
(Fig. 4E). Interestingly,
TAp63 increased Dlx3 expression
(Fig. 4E, lane 2), and this
effect was shared, although to different extents, with the EEC, SHFM and LMS
mutants (Fig. 4E; lanes 6, 7
and 8). TAp63ß did not alter Dlx3 levels
(Fig. 4E, lane 3). Remarkably,
an AEC mutant (Fig. 4E, lane 5)
and the TAp63 isoform abolished Dlx3 expression. Transfections
of Np63 isoforms, both wild-type and mutant, had no significant effect
on Dlx3 transcription (Fig.
4E, bottom panel). TAp63 and TAp63 are potent
transactivators (Barbieri and Pietenpol,
2006) and, in our in vitro studies, both isoforms were able to
transactivate Dlx3 at similar levels. However, whereas TAp63
induction was mediated through either of the two p63 sites in the
Dlx3 promoter, TAp63 required an intact regulatory region.
Surprisingly, a slightly different outcome was obtained for the endogenous
Dlx3 regulation, where upregulation of Dlx3 was detected
with TAp63 and a complete downregulation was found when expressing
TAp63 in Soas-2 cells. To determine if these results could be
attributed to cell context, the experiments were also performed in HaCaT
cells. In these cells, overexpression of TAp63 showed upregulation of
endogenous Dlx3, whereas TAp63 once again caused a complete
downregulation of Dlx3 expression (Fig.
4F). These differences might be attributed to the specific
p63RE-CCAAT-box chromatin architecture. An important feature of the
Dlx3 promoter is that the overlapping p63-binding sites are in close
proximity to CCAAT box that binds NF-Y in keratinocytes
(Park and Morasso, 1999). NF-Y
is a general promoter organizer that presets chromatin structure locally. A
recent report shows that p63 regulates the transcription of the
hsp70 gene through interactions with NF-Y
(Wu et al., 2005). Although we
have not determined the significance of NF-Y and p63 interactions on
Dlx3 transcriptional regulation, it might be proposed that there is a
dual role for the overlapping p63-binding sites, and that Dlx3 will
be transcriptionally active or repressed depending on the specific p63 isoform
bound to the promoter, on which of the p63 sites is occupied and on
interactions with NF-Y CCAAT binding factor.
Dlx3 and p63 both function as part of a complex series of cascades that
ultimately lead to the formation of ectoderm-derived organs. Unraveling the
function of each protein at specific times of embryonic development will prove
to be complex because of the differential expression of the p63 isoforms in
distinct tissues (Nylander et al.,
2002) and the cross-regulation with other developmentally relevant
signaling pathways [i.e. FGF, BMP, and Notch
(Laurikkala et al., 2006;
Nguyen et al., 2006)]. The
characterization of p63 target genes promises to improve our knowledge of the
signaling cascades that are directly involved in normal ectodermal
development. In summary, our study proves a functional relationship between
p63 and Dlx3, with Dlx3 demonstrated to be a direct target of p63.
The findings also provide evidence that the misregulation of Dlx3 is
involved in the pathogenesis of p63 molecular lesions in AEC.
ACKNOWLEDGMENTS
We thank S. J. Stimpson, Y. Rivera, S. Bertuzzi, T. Lozito, A. Pollice, K. King and G. Caretti for helpful comments; H. van Bokhoven, E. Candi and G. Melino for their gifts of plasmids; M. Simon for NHEK; G. Boekhoff-Falk for the anti-distalless antibody; and D. Roop for providing the Bmdr2 p63-KO mouse line. The research was supported by the Intramural Research Program of the NIAMS of the National Institutes of Health; Telethon (GGP05056) to L.G.; Telethon (GGP030326), AIRC and MIUR to G.L.M.; CIB to V.C.; and G.R.M. is supported by a career award from Fondazione Telethon (TCP99003) and Fondazione San Paolo, Italy.
Footnotes
REFERENCES
Acampora, D., Merlo, G. R., Paleari, L., Zerega, B., Postiglione, M. P., Mantero, S., Bober, E., Barbieri, O., Simeone, A. and Levi, G. (1999). Craniofacial, vestibular and bone defects in mice lacking the Distal-less-related gene Dlx5. Development 126,3795 -3809.[Abstract]
Barbieri, C. E. and Pietenpol, J. A. (2006). P63 and epithelial biology. Exp. Cell Res. 312,695 -706.[CrossRef][Medline]
Calabrò, V., Mansueto, G., Parisi, T., Vivo, M.,
Calogero, R. A. and La Mantia, G. (2002). The human MDM2
oncoprotein increases the transcriptional activity and the protein level of
the p53 homolog p63. J. Biol. Chem.
277,2674
-2681.
Caretti, G., Di Padova, M., Micales, B., Lyons, G. E. and
Sartorelli, V. (2004). The Polycomb Ezh2 methyltransferase
regulates muscle gene expression and skeletal muscle differentiation.
Genes Dev. 18,2627
-2638.
Ghioni, P., Bolognese, F., Duijf, P., Van Bokhoven, H.,
Mantovani, R. and Guerrini, L. (2002). Complex
transcriptional effects of p63 isoforms: identification of novel activation
and repression domains. Mol. Cell. Biol.
22,8659
-8668.
Hassan, M. Q., Javed, A., Morasso, M. I., Karlin, J., Montecino,
M., van Wijnen, A. J., Stein, G. S., Stein, J. L. and Lian, J. B.
(2004). Distal-less 3 (Dlx3) is expressed in bone: temporal
recruitment of Msx2, Dlx3 and Dlx5 homeodomain proteins to chromatin for
transcriptional regulation of osteoblast growth and differentiation.
Mol. Cell. Biol. 24,9248
-9261.
King, K. E., Ponnamperuma, R. M., Yamashita, T., Tokino, T., Lee, L. A., Young, M. F. and Weinberg, W. (2003). DeltaNp63alpha functions as both a positive and a negative transcriptional regulator and blocks in vitro differentiation of murine keratinocytes. Oncogene 22,3635 -3644.[CrossRef][Medline]
King, K. E., Ponnamperuma, R. M., Gerdes, M. J., Tokino, T.,
Yamashita, T., Baker, C. C. and Weinberg, W. C. (2006).
Unique domain functions of p63 isotypes that differentially regulate distinct
aspects of epidermal homeostasis. Carcinogenesis
27, 53-63.
Koster, M. I. and Roop, D. R. (2004). Transgenic mouse models provide new insights into the role of p63 in epidermal development. Cell Cycle 3, 411-413.[Medline]
Koster, M. I., Kim, S., Mills, A., DeMayo, F. J. and Roop, D.
R. (2004). p63 is the molecular switch for initiation of an
epithelial stratification program. Genes Dev.
18,126
-131.
Laurikkala, J., Mikkola, M. L., James, M., Tummers, M., Mills,
A. A. and Thesleff, I. (2006). P63 regulates multiple
signaling pathways required for ectodermal organogenesis and differentiation.
Development 133,1553
-1563.
McGrath, J. A., Duijf, P. H. G., Doetsch, V., Irvine, A. D., de
Wall, R., Vanmolkot, K. R. J., Wessagowit, V., Kelly, A., Atherton, D. J.,
Griffith, W. A. D. et al. (2001). Hay-Wells syndrome is
caused by heterozygous missense mutations in the SAM domain of p63.
Hum. Mol. Genet. 10,1
-9.
Merlo, G.R, Beverdam, A. and Levi, G. (2003) Dlx genes in craniofacial and limb morphogenesis. In: Murine Homeobox Gene Control of Embryonic Patterning and Organogenesis (ed. T. Lufkin). Adv. Develop. Biol. Biochem. 13,107 -132.
Mills, A. A., Zheng, B., Wang, X. J., Vogel, H., Roop, R. and Bradley, A. (1999). p63 is a p53 homologue required for limb and epidermal morphogenesis. Nature 398,708 -713.[CrossRef][Medline]
Morasso, M. I. and Radoja, N. (2005). Dlx genes, p63, and ectodermal dysplasias. Birth Defect Res. 75,163 -171.[CrossRef]
Morasso, M. I., Mahon, K. and Sargent, T. D.
(1995). A Xenopus distal-less gene in transgenic mice: conserved
regulation of distal limb epidermis and other sites of epithelial-mesenchymal
interaction. Proc. Natl. Acad. Sci. USA
92,3968
-3972.
Morasso, M. I., Markova, N. G. and Sargent, T. D.
(1996). Regulation of epidermal differentiation by a
Distal-less homeodomain gene. J. Cell Biol.
135,1879
-1887.
Morasso, M. I., Grinberg, A., Robinson, G., Sargent, T. D. and
Mahon, K. A. (1999). Placental failure in mice lacking the
homeobox gene Dlx3. Proc. Natl. Acad. Sci. USA
96,162
-167.
Nguyen, B. C., Lefort, K., Mandinova, A., Antonini, D., Devgan,
V., Della Gatta, G., Koster, M., Zhang, Z., Wang, J., Tommasi di Vignano, A.
et al. (2006). Cross-regulation between Notch and p63 in
keratinocyte commitment to differentiation. Genes Dev.
20,1028
-1042.
Nylander, K., Vojtesek, B., Nenutil, R., Lindgren, B., Roos, G., Zhanxiang, W., Sjostrom, B., Dahlqvist, A. and Coates, P. J. (2002). Differential expression of p63 isoforms in normal tissues and neoplastic cells. J. Pathol. 198,417 -427.[CrossRef][Medline]
Osada, M., Park, H. L., Nagakawa, Y., Yamashita, K., Fomenkov,
A., Kim, M. S., Wu, G., Nomoto, S., Trink, B. and Sidransky, D.
(2005). Differential recognition of response elements determines
target gene specificity for p53 and p63. Mol. Cell.
Biol. 25,6077
-6089.
Panganiban, G. and Rubenstein, J. L. R. (2002).
Developmental functions of the Distal-less/Dlx homeobox genes.
Development 129,4371
-4386.
Park, G. T. and Morasso, M. I. (1999).
Regulation of the Dlx3 homeobox gene upon differentiation of mouse
keratinocytes. J. Biol. Chem.
274,26599
-26608.
Pispa, J. and Thesleff, I. (2003). Mechanisms of ectodermal organogenesis. Dev. Biol. 262,195 -205.[CrossRef][Medline]
Price, J. A., Bowden, D. W., Wright, J. T., Pettenati, M. J. and
Hart, T. C. (1998). Identification of a mutation in DLX3
associated with tricho-dento-osseous (TDO) syndrome. Hum. Mol.
Genet. 7,563
-569.
Priolo, M. and Lagana, C. (2001). Ectodermal
dysplasias: a new clinical-genetic classification. J. Med.
Genet. 38,579
-585.
Qiao, F. and Bowie, J. U. (2005). The many faces of SAM. Sci. STKE 286, re7:1 -10.
van Bokhoven, H. and Brunner, H. G. (2002). Splitting p63. Am. J. Hum. Genet. 71, 1-13.[CrossRef][Medline]
van Bokhoven, H., Hamel, B. C., Bamshad, M., Sangiorgi, E., Gurrieri, F., Duijf, P., Vanmolkot, K., van Beusekom, E., van Beersum, S., Celli, J. et al. (2001). P63 Gene mutations in EEC syndrome, limb-mammary syndrome, and isolated split hand-split foot malformation suggest a genotype-phenotype correlation. Am. J. Hum. Genet. 69,481 -492.[CrossRef][Medline]
Wu, G., Osada, M., Guo, Z., Fomenkov, A., Begum, S., Zhao, M.,
Upadhyay, S., Xing, M., Wu, F., Moon, C. et al. (2005).
DeltaNp63alpha up-regulates the Hsp70 gene in human cancer. Cancer
Res. 65,758
-766.
Yang, A., Kaghad, M., Wang, Y., Gillett, E., Fleming, M. D., Dotsch, V., Andrews, N. C., Caput, D. and McKeon, F. (1998). p63, a p53 homolog at 3q27-29, encodes multiple products with transactivating, death-inducing, and dominant-negative activities. Mol. Cell. 2,305 -316.[CrossRef][Medline]
Yang, A., Schweitzer, R., Sun, D., Kaghad, M., Walker, R. T., Bronson, C., Tabin, A., Sharpe, D., Caput, D., Crum, C. and McKeon, F. (1999). p63 is essential for regenerative proliferation in limb, craniofacial and epithelial development. Nature 398,714 -718.[CrossRef][Medline]
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