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Tiêu đề Roles of AP-2 transcription factors in the regulation of cartilage and skeletal development
Tác giả Ann-Kathrin Wenke, Anja K. Bosserhoff
Trường học Institute of Pathology, University of Regensburg
Chuyên ngành Developmental biology
Thể loại Review article
Năm xuất bản 2009
Thành phố Regensburg
Định dạng
Số trang 9
Dung lượng 266,21 KB

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Roles of AP-2 transcription factors in the regulation ofcartilage and skeletal development Ann-Kathrin Wenke and Anja K.. AP-2a, AP-2b and AP-2c show partially overlap-ping expression pa

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Roles of AP-2 transcription factors in the regulation of

cartilage and skeletal development

Ann-Kathrin Wenke and Anja K Bosserhoff

Institute of Pathology, University of Regensburg, Germany

The AP-2 family

AP-2a was first identified by its ability to bind to

enhan-cer regions of SV40 and human metallothionein IIA [1]

The AP-2 family of transcription factors is composed of

five members: AP-2a, AP-2b, AP-2c, AP-2d, and AP-2e

[2–7], described for humans and mice Orthologs of

some AP-2s have also been found in frogs and fish, and

homologs occur in invertebrates All AP-2s have a

highly conserved basic helix–span–helix DNA-binding

and dimerization domain at their C-terminus, and a less

conserved proline-rich and glutamine-rich

transactiva-tion domain at their N-terminus [8–10] Most isoforms

also have a PY-motif (XPPXY) in the N-terminal

trans-activation domain that is important for their role as

transcriptional activators [9] The AP-2 factors form

homodimers and heterodimers for their transcriptional

activity A multiple alignment of all five human AP-2s,

illustrating their domain structure, is shown in Fig 1

A detailed and extensive overview of the AP-2 family

is given in the review of Eckert et al., [11] which also contains a schematic illustration of the AP-2 structure

Expression patterns of AP-2 molecules and functional implications

The expression and function of AP-2 isoforms have been systematically analyzed during murine embryo-genesis and in studies of the corresponding knockout mice

AP-2a, AP-2b and AP-2c show partially overlap-ping expression patterns in neural crest cells (NCCs), the peripheral nervous system, the facial mesenchyme, the limbs, various epithelia of the developing embryo,

Keywords

AP-2; cartilage; chondrogenesis; limb;

transcriptional regulation

Correspondence

A.-K Bosserhoff, Institute of Pathology,

University of Regensburg,

Franz-Josef-Strauss-Allee 11, D-93053

Regensburg, Germany

Fax: +49 941 944 6602

Tel: +49 941 944 6705

E-mail: anja.bosserhoff@klinik.uni-regens

burg.de

(Received 12 October 2009, revised 13

November 2009, accepted 20 November

2009)

doi:10.1111/j.1742-4658.2009.07509.x

During embryogenesis, most of the mammalian skeletal system is preformed

as cartilaginous structures that ossify later The different stages of cartilage and skeletal development are well described, and several molecular factors are known to influence the events of this enchondral ossification, especially transcription factors Members of the AP-2 family of transcription factors play important roles in several cellular processes, such as apoptosis, migra-tion and differentiamigra-tion Studies with knockout mice demonstrate that a main function of AP-2s is the suppression of terminal differentiation during embryonic development Additionally, the specific role of these molecules as regulators during chondrogenesis has been characterized This review gives

an overview of AP-2s, and discusses the recent findings on the AP-2 family,

in particular AP-2a, AP-2b, and AP-2e, as regulators of cartilage and skeletal development

Abbreviations

NCC, neural crest cell; RA, retinoic acid; ZPA, zone of polarizing activity.

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and the extraembryonic trophectoderm [4,12,13] In

contrast to the other AP-2s, AP-2d is specifically

expressed in the central nervous system, retina, and

developing heart [6] AP-2e expression has been

detected in the developing olfactory bulb, neural

tis-sue, especially the midbrain and hindbrain [7,14], and hypertrophic chondrocytes during chondrogenesis [15] Winger et al [16] analyzed the expression of all five mouse AP-2 family members in the unfertilized oocyte and from zygote formation to the blastocyst

Alpha -MLWKLTDNIKYEDC-EDRHDGTSNGTARLPQLGTVGQSPYTSAPPLSHT

Beta MHSPPRDQAAIMLWKLVENVKYEDIYEDRHDGVPSHSSRLSQLGSVSQGPYSSAPPLSHT

Gamma -MLWKITDNVKYEEDCEDRHDGSSNGNPRVPHLSSAGQHLYSPAPPLSHT

Epsilon -MLVHTYSAME -RPDGLG-AAAGGARLSSLPQAAYGPAPPLCHT

Delta -MSTTFPGLVHDAEIRHDGSNSYRLMQLGCLESVANSTVAYSSSSPLTYS

* : : : * :.** ::

Alpha PNA DFQPP-YFPPPY QPI-YPQSQDP -YSHVN-DPYS LNPLHAQPQP Q Beta PSS DFQPP-YFPPPY QPLPYHQSQDP -YSHVN-DPYS LNPLHQ-PQ -Q Gamma GVA EYQPPPYFPPPY QQLAYSQSADP -YSHLG-EAYAAAINPLHQPAPTGSQ Epsilon

PAATAAAEFQPP-YFPPPYPQPPLPYGQAPDAAAAFPHLAGDPYGG-LAPLAQPQPP -Delta TTG -TEFASP-YFSTNHQYTPL-HHQSFHYEFQHSHPAVTPDAYSLNSLHHSQQYYQQ :: * ** : : : *: * : *

Alpha HPGWPGQRQ -SQESGLLHTHRGLPHQLSG-LDP -RRDY -RRHEDLLHGP-HA Beta HPWGQRQRQEVGSEAGSLLPQPRAALPQLSG-LDP -RRDYHSVRRPDVLLHSAHHG Gamma QQAWPGRQSQEGAGLPSHHGRPAGLLPHLSG-LEAGAVSARRDAY RRSDLLLPHAHAL Epsilon QAAWAAPRAAARAHEE PPGLLAPPARALG-LDP -RRDYA TAVPRLLHGLADG Delta IHHGEPTDFINLHNARALKSSCLDEQRRELGCLDAYR -RHDLS LMSHGSQYGMHPD : * *: *:*

Alpha LSSGLGD-LSIHSLPH AIEEVPHVEDP -GINIPDQT-VIKKGPVSLSKSNSNAVSA Beta LDAGMGDSLSLHGLGHP-GMEDVQSVEDANNSGMNLLDQS-VIKKVPVPP -KSVTS Gamma DAAGLAENLGLHDMPH QMDEVQNVDDQ -HLLLHDQT-VIRKGPISMT KNPLN Epsilon AHGLADAPLGLPGLAAAPGLEDLQAMDEP -GMSLLDQS-VIKKVPIPSK -ASSLSA Delta

*.: ::: : : : * **::

Alpha IPINKDNLFGGV-VNPNEVFCSVPGRLSLLSSTSKYKVTVAEVQRRLSPPECLNASLLGG Beta LMMNKDGFLGGMSVNTGEVFCSVPGRLSLLSSTSKYKVTVGEVQRRLSPPECLNASLLGG Gamma LPCQKE LVGAVMNPTEVFCSVPGRLSLLSSTSKYKVTVAEVQRRLSPPECLNASLLGG Epsilon LSLAKDS-LVGGITNPGEVFCSVPGRLSLLSSTSKYKVTVGEVQRRLSPPECLNASLLGG Delta -GTCVVNPTDLFCSVPGRLSLLSSTSKYKVTIAEVKRRLSPPECLNASLLGG * ::********************:.**:****************

Alpha VLRRAKSKNGGRSLREKLDKIGLNLPAGRRKAANVTLLTSLVEGEAVHLARDFGYVCETE Beta VLRRAKSKNGGRSLRERLEKIGLNLPAGRRKAANVTLLTSLVEGEAVHLARDFGYICETE Gamma VLRRAKSKNGGRSLREKLDKIGLNLPAGRRKAAHVTLLTSLVEGEAVHLARDFAYVCEAE Epsilon VLRRAKSKNGGRCLRERLEKIGLNLPAGRRKAANVTLLTSLVEGEAVHLARDFGYVCETE Delta ILRRAKSKNGGRCLREKLDRLGLNLPAGRRKAANVTLLTSLVEGEALHLARDFGYTCETE :***********.***:*:::************:************:******.* **:*

Alpha FPAKAVAEFLNRQHSD-PNEQVTRKNMLLATKQICKEFTDLLAQDRSPLGNSRPNPILEP Beta FPAKAVSEYLNRQHTD-PSDLHSRKNMLLATKQLCKEFTDLLAQDRTPIGNSRPSPILEP Gamma FPSKPVAEYLTRPHLGGRNEMAARKNMLLAAQQLCKEFTELLSQDRTPHGTSRLAPVLET Epsilon FPAKAAAEYLCRQHAD-PGELHSRKSMLLAAKQICKEFADLMAQDRSPLGNSRPALILEP Delta FPAKAVGEHLARQHME-QKEQTARKKMILATKQICKEFQDLLSQDRSPLGSSRPTPILDL **:* *.* * * : :**.*:**::*:**** :*::***:* *.** :*:

Alpha GIQSCLTHFNLISHGFGSPAVCAAVTALQNYLTEALKAMDKMYLS -NNP-NSHTDN Beta GIQSCLTHFSLITHGFGAPAICAALTALQNYLTEALKGMDKMFLN -NTTTNRHTSG Gamma NIQNCLSHFSLITHGFGSQAICAAVSALQNYIKEALIVIDKSYMN -PGD-QSPADS Epsilon GVQSCLTHFSLITHGFGGPAICAALTAFQNYLLESLKGLDKMFLS -SVG-SGHGET Delta DIQRHLTHFSLITHGFGTPAICAALSTFQTVLSEMLNYLEKHTTHKNGGAADSGQGHANS .:* *:**.**:**** *:***::::* : * * ::*

Alpha N AKSSDKEEKHRK -

Beta EGP-GSKTGDKEEKHRK -

Gamma N -KTLEKMEKHRK -

Epsilon K -ASEKDAKHRK -

Delta EKAPLRKTSEAAVKEGKTEKTD : : : * *

Fig 1 Multiple alignment of AP-2a, AP-2b, AP-2c, AP-2d, and AP-2e The proline-rich and glutamine-rich N-terminus, which is important for transactivation, is shown in yellow, and contains the PY-motif (green) The helix–span–helix domain at the C-terminus shown in blue medi-ates dimerization and, together with the basic domain, (red) DNA-binding ‘*’, amino acids that are identical in all sequences in the align-ment; ‘:’, conserved substitutions have been observed; ‘.’, semiconserved substitutions.

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stage of development They found that AP-2a,

AP-2b, AP-2c and AP-2e are differentially expressed

during the preimplantation period, and, with the

exception of AP-2a, also in unfertilized oocytes

Furthermore, they determined that functional

redun-dancy occurs between these proteins during at least

the preimplantation period [16]

However, gene knockout experiments indicate that

the AP-2s perform individual and nonredundant

functions during mouse development Analyses of

AP-2a-null mice have demonstrated that AP-2a is a

fundamental regulator of mammalian craniofacial

development AP-2a knockout mice die perinatally

with craniofacial defects, thoracoabdominoschisis, and

severe skeletal defects in the head and trunk region

[17,18] Studies of earlier embryonic stages of these

mice indicate a failure of cranial neural tube closure

and defects in cranial ganglia development Another

role of AP-2a previously masked in the knockout mice

became apparent in chimeric mice composed of both

wild-type and AP-2a-null cells [19] These chimeras

reveal the major influence of AP-2a on eye

forma-tion and limb pattern formaforma-tion typified by limb

duplications

In contrast to these defects, the lack of AP-2b

leads to enhanced apoptotic cell death of renal

epi-thelial cells AP-2b knockout mice die shortly after

birth because of polycystic kidney disease and

termi-nal retermi-nal failure [20,21] The targeted deletion of

AP-2c also has severe consequences The loss of

AP-2c is already lethal in early embryogenic

develop-ment directly after implantation during gastrulation,

because AP-2c controls proliferation and

differentia-tion of extraembryonic trophectodermal cells [22,23]

So far, nothing is known about chondrogenic defects

mediated by knocking out AP-2b or AP-2c

However, all these types of grave damage after

deletion of AP-2 transcription factors demonstrate

the importance of the AP-2s for several functions

during embryonic development To date, knockout

studies concerning AP-2d or AP-2e have not been

published

Regulation of AP-2 and AP-2 target

genes

The expression of the AP-2a transcription factor is

induced by different signal-transducing agents, such as

retinoic acid (RA), cAMP, phorbol ester, UV light, and

singlet oxygen [2,24–26] RA plays an important role in

the process of chondrocyte differentiation [27] AP-2

mediates transcriptional activation in response to two

different signal transduction pathways, the phorbol

ester-activated protein kinase C pathway, or the cAMP-dependent protein kinase A pathway [28] Here, cAMP may modulate AP-2 activity by protein kinase A-induced phosphorylation of the transcription factor [29]

So far, interactions with AP-2 have been described for many proteins For example, CBP⁄ p300-interacting transactivator with ED-rich tail 2 interacts with and co-activates all isoforms of AP-2, and the interaction with AP-2a is suggested to be necessary for normal neural tube and cardiac development [30,31] The Kru¨ppel-related zinc finger protein AP-2rep (Klf12) has been characterized as a repressor of AP-2a Repression of AP-2a transcription by AP-2rep is dependent on an N-terminal PVDLS motif that interacts specifically with the corepressor CtBP1 [32,33] Recently, it was shown that the broad-complex, tramtrack and bric-a-brac domain containing protein KCTD1 directly binds to AP-2a and acts as a negative regulator for AP-2a trans-activation [34] It was also demonstrated in other studies that the nuclear protein poly(ADP-ribose) polymerase-1 interacts with the C-terminus of AP-2a and enhances its transcriptional activity in normal circumstances, whereas its enzymatic activity is used as a temporary shut-off mechanism during unfavorable conditions [35,36] Little is known about the interaction of AP-2 and its binding partners in cartilage However, at least CBP⁄ p300-interacting transactivator with ED-rich tail 2 and protein poly(ADP-ribose) polymerase-1 are expressed in this tissue [37–40] It would be interesting

to further analyze their interactions with AP-2 and the functional role of these in chondrocytes

Furthermore, it is speculated that in melanoma, where AP-2a acts as a tumor suppressor, the loss of AP-2a is caused by a failure in post-transcriptional processing of the protein [41] Additionally, it is evi-dent that AP-2 transcription factors can indirectly modulate genes by functional interactions with other transcription factors, e.g c-myc, rBP, and p53 [42–44] The formation of AP-2 homodimers and heterodimers could also be important for their regulatory activity, but no studies have been published so far

For the regulation of target gene expression, the AP-2 transcription factors bind to the palindromic recognition sequence 5¢-GCCN3GGC-3¢ or variations

of this GC-rich sequence within multiple gene promot-ers [45] AP-2s play a dual role as transcriptional acti-vators and repressors By regulating target genes with AP-2-binding sites within their promoter sequences, the AP-2 transcription factors play important roles in cellular processes, such as morphogenesis, in particular proliferation, differentiation, cell cycle regulation, and apoptosis [11,45,46] Through suppression of genes inducing terminal differentiation, apoptosis, and

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growth retardation, AP-2s play vital roles in cell

prolif-eration Besides the functions of AP-2s in physiological

processes, they have also crucial roles in pathological

processes such as tumorigenesis and genetic diseases

[47]

Most analyses of the regulation of AP-2 and the

interactions of the transcription factor with binding

partners, as well as of the regulation of target gene

expression, have been performed for AP-2a Up to

now, there have been no similar studies for the other

AP-2 isoforms

Chondrogenesis and skeletal

development

Most elements of the vertebrate skeleton are built

through enchondral ossification This is a complex

pro-cess beginning with the migration of undifferentiated

mesenchymal cells to regions determined to

differenti-ate into bone, followed by aggregation and the

forma-tion of mesenchymal condensaforma-tion [48,49] These

resting and proliferating chondrocytes produce an

extracellular matrix mainly consisting of aggrecan and

type II collagen As skeletogenesis proceeds,

proliferat-ing chondrocytes exit the cell cycle, become

hypertro-phic, express type X collagen, and reduce the

expression of type II collagen [50] Hypertrophic

chon-drocytes undergo terminal differentiation before they

finally become apoptotic Through the invasion of

blood vessels from the perichondrium, the cartilage

becomes vascularized Additionally, osteoblasts invade

the cartilage and start to replace it with mineralized

bone [48]

Many molecules and signaling cascades are

neces-sary to regulate these molecular processes of

chondro-genic and skeletal development, including transcription

factors Essential transcription factors in chondrocyte

differentiation are Sox9 and Runx2 Sox9 plays a key

role in chondrogenesis, as an inactivating mutation in

the gene encoding Sox9 leads to severe cartilage

abnor-malities called campomelic dysplasia [51,52] The effect

of a complete loss of Sox9 during chondrogenesis was

analyzed using a model of mice chimeras injected with

homozygous embryonic Sox9) ⁄ ) stem cells into

wild-type blastocysts, because Sox9 knockout mice are not

viable [53] The Sox9) ⁄ )cells were excluded from

mes-enchymal condensation and had no expression of the

chondrocytic markers type II collagen, type IX

colla-gen, type X collacolla-gen, and aggrecan Besides type II

collagen and aggrecan, Sox9 also regulates the

expres-sion of the cartilage-derived retinoic acid-sensitive

pro-tein [54,55] Sox5 and Sox6, members of the Sox

family, are also important for chondrocyte

differentia-tion, as embryos lacking Sox5 and Sox6 die at embry-onic day 16.5 and display a failure of chondrocyte progenitor cells to differentiate into hypertrophic chon-drocytes [56]

Two members of the Runx family of transcription factors, Runx2 and Runx3, are positive regulators of chondrocyte hypertrophy Runx2 is transiently expressed in prehypertrophic chondrocytes, and enforced expression of Runx2 in these cells in trans-genic mice leads to ectopic chondrocyte hypertrophy [57] Mice lacking both Runx2 and Runx3 do not have hypertrophic chondrocytes or type X collagen-express-ing cells, showcollagen-express-ing that both Runx2 and Runx3 are important regulators for hypertrophic development of chondrocytes [58] Alongside the important function for chondrogenesis, Runx2 is also a key regulator for osteoblast differentiation In particular, Runx2 is expressed in cells prefiguring the vertebrate skeleton as early as embryonic day 10.5 [59] Runx2 regulates many genes that determine the osteoblast phenotype,

as the forced expression of Runx2 in nonosteoblast cells is sufficient to induce the osteoblast-specific gene osteocalcin[60] The inactivation of both Runx2 alleles

in mice results in a lack of osteoblasts throughout the skeleton [61,62] It has also been shown that deletions resulting in the heterozygous loss of runx2 cause cleid-ocranial dysplasia [63]

Role of AP-2a, AP-2b and AP-2e in chondrogenesis and skeletal development

In addition to Sox and Runx transcription factors, members of the AP-2 family also have important func-tions in chondrogenesis and development of the verte-brate skeleton during embryogenesis Especially for AP-2a, but also for AP-2b and AP-2e, a role as a reg-ulator of cartilage differentiation has been shown [64–69] The functional and important roles of AP-2 transcription factors during chondrogenesis are illus-trated in Fig 2

AP-2a is expressed in the growth plate and in articu-lar cartilage, and has been described as a negative regulator of chondrocyte differentiation [64] The expression of cartilage-derived retinoic acid-sensitive protein and type II collagen is negatively correlated with AP-2a expression, and AP-2a thus acts as a sup-pressor of these two cartilage matrix genes during car-tilage differentiation [64–66] (Fig 2) High expression levels of AP-2a in chondroprogenitor cells maintain these cells in an early differentiation phenotype and inhibit the transition to differentiated chondrocytes The induction of Sox5 and Sox6 as well as that of chondrocytic matrix genes such as type II collagen,

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aggrecan and type X collagen are also delayed by

AP-2a [64,67]

Reports on AP-2a knockout mice clearly indicate

the importance of this transcription factor in

regulat-ing bone and cartilage development durregulat-ing

embryogen-esis, because of the severe skeletal defects in growth

and the development of face and limbs [17–19]

Don-ner et al tried to link the expression of AP-2a in these

tissues to upstream signaling pathways They assessed

the organization of a cis-regulatory region within the

fifth intron specific for directing AP-2a expression to

the developing frontal nasal process and limb bud

mes-enchyme, which they had previously identified in

trans-genic mice [70,71] The results demonstrate that a

STAT binding site is required for robust AP-2a

expres-sion in the face and limbs In a follow-up study, they

found that this conserved cis-acting sequence serves to

maintain a level of AP-2a expression that limits the

size of the hand plate and the associated number of

digit primordia [72]

AP-2 function was also analyzed in other species

A similar role for AP-2a as a regulator for face and

limb bud development was described in chickens AP-2

expression is completely downregulated after treatment

of the chick face with RA, and this is accompanied by

an increase in apoptosis [73] The authors of this study

ascribe the regulation of outgrowth of limb buds and

patterning of the digits to the chicken AP-2

The role of AP-2a was further studied in zebrafish

It was confirmed that AP-2a is an essential regulator

of the development of neural crest derivates, including embryonic cartilage and neurons, as well as pigmented cells [74–76] Knight et al [77] demonstrated essential functions for zebrafish AP-2a (tfap2a) and also AP-2b (tfap2b) in the development of the facial ectoderm, and for signals from this epithelium that induce skeletogen-esis in NCCs Zebrafish embryos lacking both tfap2a and tfap2b have defects in epidermal cell survival and deficient NCC-derived cartilage The authors propose that AP-2s have two distinct functions in cranial NCCs: they play an early cell-autonomous role in cell specification and survival, and a later nonautonomous role as regulators of ectodermal signals that induce skeletogenesis [77]

Luo et al [78] characterized Inca (induced in the neural crest by AP-2) as a target gene upregulated by AP-2a in Xenopus embryos Knockdown experiments for Inca in frog and fish revealed essential functions in

a subset of NCCs that form craniofacial cartilage Cells deficient for Inca show normal migration but fail

to condense into skeletal primordia This is an interest-ing aspect, as, for murine embryonic development, AP-2a is described as a suppressor of cartilage differ-entiation, maintaining cells in an early differentiated phenotype

For AP-2b, expression in murine limbs has also been demonstrated AP-2b is expressed in the zone of polar-izing activity (ZPA), the signaling center of the devel-oping vertebrate limb [68] A microarray approach comparing gene expression in the ZPA with that in the

Sox9

AP-2ε

Undifferentiated

mesenchymal cells

Differentiated

chondrocytes

Hypertrophic

chondrocytes

Condensed

mesenchymal cells

Sox9

Sox9

Sox5

Sox6

AP-2α

Runx2 Runx3 Runx2

Runx2

Fig 2 Functional role of AP-2a and AP-2e

in chondrogenesis Overview of the differen-tiation stages during chondrogenesis and the involvement of transcription factors (henatoxylin and eosin-stained section of an embryonic cartilaginous limb).

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rest of the limb showed that AP-2b expression is

increased in the ZPA

The fifth member of the AP-2 family, AP-2e, is

expressed in human articular cartilage, where it has

been shown to be a regulator of integrin a10

expres-sion [15] Recently, it was reported that the

transcrip-tion factor Sox9 induces AP-2e expression in the

hypertrophic stage of chondrocytic

differentia-tion through direct binding to the AP-2e promoter [69]

(Fig 2) Additionally, osteoarthritis chondrocytes show

increased expression of AP-2e as compared with

differentiated chondrocytes [69] Further studies are

required to identify AP-2e target genes other than

integrin a10, to clarify the role of AP-2e in

chon-drocyte differentiation and in the development of

osteoarthritis

Role of AP-2 in chondrocytic diseases

A role for AP-2s as regulators has been shown for

sev-eral chondrogenic diseases For example, mutations in

tfap2a are known to cause branchio-oculo-facial

syn-drome [79] The characteristic craniofacial features of

this disease are dolichocephaly, malformed pinnae,

thick nasal tip, and cleft lip Moreover, it has been

reported that branchio-oculo-facial syndrome has

over-lapping features, such as orofacial clefting and

occa-sional lip pits, with Van der Woude syndrome, in

which disruption of an AP-2a-binding site within an

interferon regulatory factor 6 enhancer is strongly

associated with cleft lip [80] Recently, it has been

demonstrated that AP-2e is overexpressed in

osteoar-thritic chondrocytes, but the exact function of AP-2e

in osteoarthritic development of cartilage is still

unknown [69]

Conclusions

AP-2 proteins, especially AP-2a and AP-2e, are

impor-tant for chondrogenic and skeletal development Many

studies on AP-2a have been performed, analyzing the

role of this transcription factor as a main regulator

of facial and limb development in embryogenesis

Further analyses are required to clarify the regulatory

mechanisms during early chondrocytic differentiation,

because it is still unknown how AP-2a itself is

upregu-lated in chondroprogenitor cells The molecular

rele-vance of AP-2e in hypertrophic cartilage and in the

development of osteoarthritis also still has to be

ana-lyzed in detail It is necessary to obtain more insights

into the transcriptional regulation of AP-2s, to

under-stand the complex story of AP-2s during embryonic

development

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