RESUMO
The adenomatous polyposis coli (APC) tumour-suppressor protein controls the Wnt signalling pathway by forming a complex with glycogen synthase kinase 3beta (GSK-3beta), axin/conductin and betacatenin. Complex formation induces the rapid degradation of betacatenin. In colon carcinoma cells, loss of APC leads to the accumulation of betacatenin in the nucleus, where it binds to and activates the Tcf-4 transcription factor (reviewed in [1] [2]). Here, we report the identification and genomic structure of APC homologues. Mammalian APC2, which closely resembles APC in overall domain structure, was functionally analyzed and shown to contain two SAMP domains, both of which are required for binding to conductin. Like APC, APC2 regulates the formation of active betacatenin-Tcf complexes, as demonstrated using transient transcriptional activation assays in APC -/- colon carcinoma cells. Human APC2 maps to chromosome 19p13.3. APC and APC2 may therefore have comparable functions in development and cancer.
Assuntos
Polipose Adenomatosa do Colo/genética , Proteínas do Citoesqueleto/genética , Genes APC , Neuropeptídeos/genética , Serina Endopeptidases/genética , Sequência de Aminoácidos , Animais , Proteínas do Citoesqueleto/química , Humanos , Camundongos , Dados de Sequência Molecular , Neuropeptídeos/química , Homologia de Sequência de Aminoácidos , Serina Endopeptidases/químicaRESUMO
Early embryonic development in Xenopus laevis is characterized by transcriptional repression which is relieved at the midblastula stage (MBT). Here we show that the relative abundance of TATA-binding protein (TBP) increases robustly at the MBT and that the mechanism underlying this increase is translation of maternally stored TBP RNA. We show that TBP is rate-limiting in egg extract under conditions that titrate nucleosome assembly. Precocious translation of TBP mRNA in Xenopus embryos facilitates transcription before the MBT, without requiring TBP to be prebound to the promoter before injection. This effect is transient in the absence of chromatin titration and is sustained when chromatin is titrated. These data show that translational regulation of TBP RNA contributes to limitations on the transcriptional capacity before the MBT. Second, we examined the ability of trans-acting factors to contribute to promoter activity before the MBT. Deletion of cis-acting elements does not affect histone H2B transcription in egg extract, a finding indicative of limited trans-activation. Moreover, in the context of the intact promoter, neither the transcriptional activator Oct-1, nor TBP, nor TFIID enable transcriptional activation in vitro. HeLa cell extract, however, reconstitutes activated transcription in mixed extracts. These data suggest a deficiency in egg extract cofactors required for activated transcription. We show that the capacity for activated H2B transcription is gradually acquired at the early gastrula transition. This transition occurs well after the blastula stage when the basal transcription machinery can first be complemented with TBP.
Assuntos
Proteínas de Ligação a DNA/genética , Regulação da Expressão Gênica no Desenvolvimento , Biossíntese de Proteínas , Fatores de Transcrição/genética , Ativação Transcricional , Xenopus laevis/embriologia , Animais , Blastocisto/metabolismo , Proteínas de Ligação a DNA/metabolismo , Óvulo , RNA Mensageiro/metabolismo , Proteína de Ligação a TATA-Box , Fatores de Transcrição/metabolismoRESUMO
Tcf transcription factors interact with beta-catenin and Armadillo to mediate Wnt/Wingless signaling. We now report the characterization of genes encoding two murine members of the Tcf family, mTcf-3 and mTcf-4. mTcf-3 mRNA is ubiquitously present in embryonic day 6.5 (E6.5) mouse embryos but gradually disappears over the next 3 to 4 days. mTcf-4 expression occurs first at E10.5 and is restricted to di- and mesencephalon and the intestinal epithelium during embryogenesis. The mTcf-3 and mTcf-4 proteins bind a canonical Tcf DNA motif and can complex with the transcriptional coactivator beta-catenin. Overexpression of Wnt-1 in a mammary epithelial cell line leads to the formation of a nuclear complex between beta-catenin and Tcf proteins and to Tcf reporter gene transcription. These data demonstrate a direct link between Wnt stimulation and beta-catenin/Tcf transcriptional activation and imply a role for mTcf-3 and -4 in early Wnt-driven developmental decisions in the mouse embryo.
Assuntos
Proteínas do Citoesqueleto/metabolismo , Proteínas HMGB , Proteínas Proto-Oncogênicas/metabolismo , Transdução de Sinais , Transativadores , Fatores de Transcrição/metabolismo , Proteínas de Peixe-Zebra , Células 3T3 , Sequência de Aminoácidos , Animais , Linhagem Celular , Núcleo Celular/metabolismo , Galinhas , Clonagem Molecular , Desenvolvimento Embrionário e Fetal , Expressão Gênica , Genes Reporter , Humanos , Luciferases/genética , Camundongos , Dados de Sequência Molecular , Células PC12 , Proteínas Proto-Oncogênicas/biossíntese , Proteínas Proto-Oncogênicas/genética , Ratos , Fatores de Transcrição TCF , Proteína 1 Semelhante ao Fator 7 de Transcrição , Proteína 2 Semelhante ao Fator 7 de Transcrição , Fatores de Transcrição/biossíntese , Ativação Transcricional , Proteínas Wnt , Proteína Wnt1 , beta CateninaRESUMO
The Xenopus homolog of the proto-oncogene wnt-1 (int-1) is transiently expressed during neurula and tailbud stages of early development. To determine the mechanisms involved in the transcriptional regulation of the Xwnt-1 gene, we isolated Xwnt-1 genomic sequences. The promoter activity of the 5' flanking region of the gene was analysed by microinjection of chimeric luciferase reporter constructs into embryos. It is shown that the proximal 220 bp of the Xwnt-1 promoter is able to confer activation of the reporter gene at the early neurula stage. DNaseI footprinting analysis of the proximal promoter region with nuclear protein extracts from neurula stage embryos revealed five protein binding regions. Deletion and mutation analysis shows that one of these five protein binding regions is essential for promoter activity. Gel shift experiments indicate that a sequence element resembling the GT-I and GT-II motifs of the SV40 enhancer is important for the Xwnt-1 promoter activity in vivo.
Assuntos
Regiões Promotoras Genéticas/genética , Proteínas Proto-Oncogênicas/genética , Xenopus laevis/genética , Proteínas de Peixe-Zebra , Sequência de Aminoácidos , Animais , Sequência de Bases , Quimera/genética , Mapeamento Cromossômico , DNA/genética , Análise Mutacional de DNA , Desenvolvimento Embrionário e Fetal/genética , Feminino , Deleção de Genes , Regulação da Expressão Gênica/genética , Genes Reporter/genética , Luciferases/genética , Dados de Sequência Molecular , Ligação Proteica/genética , Transcrição Gênica/genética , Proteínas Wnt , Proteína Wnt1 , Proteínas de XenopusRESUMO
1. We have investigated the origin of proteolytic activity which causes degradation of histones in chromatin isolated from Xenopus liver and the rat liver at neutral pH. Polyacrylamide disc gel electrophoresis was used for detection of proteolytic products of histones. 2. No proteolytic degradation of histones occurs in chromatin isolated from Xenopus erythrocytes and rat liver according to our procedure even after prolonged incubation at pH 8.0 and pH 5.0. However with chromatin isolated from Xenopus liver a high level of histone degradation is observed under similar conditions. 3. Mixing isolated nuclei from Xenopus erythrocytes with a crude cytoplasmic fraction from Xenopus liver causes histone proteolysis in isolated chromatin at pH 8.0. In similar experiments with corresponding fractions from rat liver histone proteolysis can be introduced only after repeated freezing and thawing of the cytoplasmic fraction. 4. A purified lysosomal preparation from rat liver causes a similar type of histone degradation upon incubation with chromatin from Xenopus erythrocytes and rat liver. 5. The neutral proteolytic activity that can be introduced in isolated chromatin by a crude cytoplasmic fraction and by a purified lysosomal erythrocytes and rat liver. 5. The neutral proteolytic activity that can be introduced in isolated chromatin by a crude cytoplasmic fraction and by a purified lysosomal fraction from rat liver is inhibited by sodium bisulphite. 6. We conclude that the neutral proteolytic activity which causes degradation of histones in isolated chromatin is due to a contamination with neutral protease(s) originating from cytoplasmic organelles.
Assuntos
Fígado/enzimologia , Peptídeo Hidrolases/análise , Animais , Núcleo Celular/enzimologia , Cromatina , Citoplasma/enzimologia , Eritrócitos/enzimologia , Histonas , Concentração de Íons de Hidrogênio , Lisossomos/enzimologia , Ratos , XenopusRESUMO
Lysine-rich histones have been isolated from the terminally differentiated erythrocytes of Xenopus laevis. Three major proteins have been separated by ion-exchange chromatography. These proteins have been characterized by electrophoresis, amino acid analysis and immunochemical techniques. It is concluded that two 'typical' lysine-rich subfractions are present in Xenopus erythrocytes and, in addition, a serine-rich histone, that shares no common antigenic determinants with the other lysine-rich histones.
Assuntos
Eritrócitos/análise , Histonas , Lisina , Aminoácidos/análise , Animais , Galinhas , Testes de Fixação de Complemento , Histonas/sangue , Peso Molecular , Especificidade da Espécie , XenopusRESUMO
To study the possible role of the glucocorticoid receptor (GR) in early embryogenesis, we isolated a Xenopus glucocorticoid receptor cDNA from an embryonic stage 17 cDNA library. Overexpression of this Xenopus GR in COS cells confers the ability to transactivate a GRE-tk CAT promoter construct in a ligand dependent manner. Expression of the Xenopus GR gene at the RNA level was analyzed by Northern blot hybridization. Transcripts of 4 and 6 kb are present in oocytes. The 4 kb mRNA is abundant and is degraded together with the 6 kb mRNA during cleavage stages of early development. Between stages 17 and 24, GR messengers are extremely rare. From stage 32 onwards, both GR transcripts start to be expressed again at intermediate levels. These results provide the first evidence that expression of the GR gene is regulated during early embryonic development.
Assuntos
DNA Complementar/isolamento & purificação , Regulação da Expressão Gênica , Receptores de Glucocorticoides/genética , Xenopus laevis/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA Complementar/química , Dados de Sequência Molecular , RNA Mensageiro/genética , Xenopus laevis/embriologiaRESUMO
Using previously cloned Xenopus nucleosomal core histone genes as hybridization probes, a genomic DNA library of Xenopus laevis was screened for histone gene clusters. From over 200 histone-gene containing clones identified, 36 were selected as possibly containing H1 histone genes by hybridization to a probe derived from a sea urchin H1 histone gene. These 36 clones were further analyzed by hybrid-selected translation for the definitive presence of H1 histone genes. The genes for three different H1 histone variants were found: H1A , H1B and H1C . Mapping of the histone genes within each clone showed that at least three different gene arrangements can occur within a cluster and that the type of H1 histone variant present in a cluster may be related to the cluster type. S1-mapping experiments indicated that histone genes found in different cluster-types can be expressed in oocytes. Also, the H1 gene found in one cluster-type was expressed in at least three different cell-types: oocytes, gastrula-stage embryos, and erythroblasts.
Assuntos
Genes , Variação Genética , Histonas/genética , Transcrição Gênica , Animais , Clonagem Molecular , Enzimas de Restrição do DNA , Vetores Genéticos , Hibridização de Ácido Nucleico , Biossíntese de Proteínas , RNA Mensageiro/genética , XenopusRESUMO
Oct-1, a member of the POU family of transcription factors, is expressed at relatively high levels in ectodermal and mesodermal cell lineages during early Xenopus embryogenesis (Veenstra et al, 1995). Here we show that overexpression of Oct-1 induces programmed cell death concomitant with the loss of the posterior part of the body axis. Truncated Oct-1 variants, missing either the C-terminal or N-terminal trans-activation domain, exhibit a different capacity to cause such developmental defects. Oct-1-induced cell death is rescued in unilaterally injected embryos by non-injected cells, indicative of the non-cell autonomous character of the developmental effects of Oct-1. This was confirmed by marker gene analysis, which showed a significant decrease in brachyury expression, suggesting that Oct-1 interferes with an FGF-type signalling pathway.
Assuntos
Apoptose , Proteínas de Ligação a DNA/biossíntese , Proteínas Fetais , Proteínas com Domínio T , Fatores de Transcrição/biossíntese , Animais , Sítios de Ligação , Biomarcadores , Proteínas de Ligação a DNA/genética , Gástrula , Fator C1 de Célula Hospedeira , Morfogênese , Fator 1 de Transcrição de Octâmero , Fatores de Transcrição/genética , Ativação Transcricional , Xenopus/embriologia , Proteínas de XenopusRESUMO
Recently, we demonstrated that the Xenopus Wnt effector XTcf-3 interacts with Groucho-related transcriptional repressors (Roose et al., 1998. Nature 395, 608-612). A long form of the Groucho-related genes, XGrg-4, was shown to repress axis formation in the Xenopus embryo, whereas a short form, XGrg-5, acted as a potentiator. In this study, the temporal and spatial expression of XGrg-4 and XGrg-5 is described in Xenopus laevis embryos. Both genes are maternally expressed. In the gastrula, transcripts of both genes are present in the animal as well as the vegetal region. At later stages, XGrg-4 and XGrg-5 show specific patterns of expression in the central nervous system (CNS), cranial ganglia, eyes, otic vesicles, stomodeal-hypophyseal anlage, cement gland, head mesenchyme, branchial arches, neural crest and derivatives, somites, pronephros, pronephric duct, heart and tailbud. Differences in the expression of XGrg-4 and XGrg-5 were found in the CNS, cranial ganglia, olfactory placodes, stomodeal-pharyngeal anlage, cement gland, head mesenchyme and ectoderm.
Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Proteínas Repressoras/genética , Xenopus laevis/embriologia , Animais , Proteínas Correpressoras , Proteínas de Ligação a DNA , Proteínas de Xenopus , Xenopus laevis/genéticaRESUMO
Recent evidence that Wnts and other genes in the Wnt signaling pathway are expressed in embryonic and adult mouse lung suggests that this pathway is important for cell fate decisions and differentiation of lung cell types. We therefore examined the expression and protein distribution of several Wnt pathway components during prenatal mouse lung development using whole-mount in situ hybridization and immunohistochemistry. Between embryonic days 10.5 and 17.5 (E10.5-E17.5), beta-catenin was localized in the cytoplasm, and often also the nucleus, of the undifferentiated primordial epithelium (PE), differentiating alveolar epithelium (AE; present from E14.5 onward), and adjacent mesenchyme. Tcf1, Lef1, Tcf3, Tcf4, sFrp1, sFrp2 and sFrp4 were also expressed in the PE, AE, and adjacent mesenchyme in specific spatio-temporal patterns.
Assuntos
Proteínas do Citoesqueleto/biossíntese , Proteínas de Ligação a DNA/biossíntese , Glicoproteínas/biossíntese , Pulmão/embriologia , Transativadores , Fatores de Transcrição/biossíntese , Animais , Diferenciação Celular , Linhagem da Célula , Citoplasma/metabolismo , Fator 1-alfa Nuclear de Hepatócito , Imuno-Histoquímica , Hibridização In Situ , Peptídeos e Proteínas de Sinalização Intracelular , Fator 1 de Ligação ao Facilitador Linfoide , Camundongos , Fator 1 de Transcrição de Linfócitos T , Fatores de Tempo , beta CateninaRESUMO
The spatio-temporal expression of connexin43 in Xenopus laevis embryos was studied by in situ hybridization. Cx43 expression is first detected at stage 25 in the developing eye. In stage 32, expression was found in the margin of the lens placode, the cement gland, notochord, and in stage 37 in the branchial arches. Early limb buds show strong expression of Cx43 distally while later on expression is confined to sites of precartilage condensation.
Assuntos
Conexina 43/genética , Xenopus laevis/embriologia , Xenopus laevis/genética , Sequência de Aminoácidos , Animais , Extremidades/embriologia , Olho/embriologia , Regulação da Expressão Gênica no Desenvolvimento , Hibridização In Situ , Dados de Sequência Molecular , Notocorda/embriologia , Homologia de Sequência de Aminoácidos , Especificidade da Espécie , Xenopus/embriologia , Xenopus/genéticaRESUMO
The recent discovery that the HMG box transcription factor XTCF-3 is involved in early axis specification in Xenopus laevis (Molenaar, M., van de Wetering, M., Oosterwegel, M., Peterson-Maduro, J. Godsave, S., Korinek, V., Roose, J., Destree, O., Clevers, H., 1996. XTcf-3 transcription factor mediates beta-catenin-induced axis formation in Xenopus embryos. Cell 86, 391-399) led us to search for other members of the TCF/LEF family in this species. A newly identified HMG box factor was cloned with highest homology to human LEF-1, called XLEF-1. Unlike XTcf-3, XLef-1 is not expressed maternally, but its transcripts become detectable directly after the mid blastula transition (MBT). At later stages, both genes are expressed in the central nervous system (CNS), eyes, otic vesicles, head mesenchyme, neural crest and derivatives, branchial arches, developing heart, tailbud and limb buds. The expression pattern of Lef-1 during later stages of development is evolutionarily conserved.
Assuntos
Proteínas HMGB , Proteínas de Grupo de Alta Mobilidade/genética , Fatores de Transcrição/genética , Proteínas de Xenopus , Xenopus/genética , Sequência de Aminoácidos , Animais , Northern Blotting , Clonagem Molecular , DNA Complementar/química , DNA Complementar/genética , Embrião não Mamífero/metabolismo , Desenvolvimento Embrionário , Regulação da Expressão Gênica no Desenvolvimento , Hibridização In Situ , Fator 1 de Ligação ao Facilitador Linfoide , Dados de Sequência Molecular , RNA/análise , RNA/genética , RNA Mensageiro/análise , RNA Mensageiro/genética , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Fatores de Transcrição TCF , Fator 3 de Transcrição , Proteína 1 Semelhante ao Fator 7 de Transcrição , Xenopus/embriologiaRESUMO
The Frat1 gene was first identified as a proto-oncogene involved in progression of mouse T cell lymphomas. More recently, FRAT/GBP (GSK-3beta Binding Protein) family members have been recognized as critical components of the Wnt signal transduction pathway. In an attempt to gain more insight into the function of Frat1, we have generated Frat1-deficient mice in which most of the coding domain was replaced by a promoterless beta-galactosidase reporter gene. While the pattern of LacZ expression in Frat1(lacZ)/+ mice indicated Frat1 to be expressed in various neural and epithelial tissues, homozygous Frat1(lacZ) mice were apparently normal, healthy and fertile. Tissues of homozygous Frat1(lacZ) mice showed expression of a second mouse Frat gene, designated Frat3. The Frat1 and Frat3 proteins are structurally and functionally very similar, since both Frat1 and Frat3 are capable of inducing a secondary axis in Xenopus embryos. The overlapping expression patterns of Frat1 and Frat3 during murine embryogenesis suggest that the apparent dispensability of Frat1 for proper development may be due to the presence of a second mouse gene encoding a functional Frat protein.
Assuntos
Proteínas de Transporte , Proteínas de Neoplasias , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas/metabolismo , Proteínas de Xenopus , Proteínas Adaptadoras de Transdução de Sinal , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Embrião não Mamífero , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Teste de Complementação Genética , Homozigoto , Peptídeos e Proteínas de Sinalização Intracelular , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Mutantes , Dados de Sequência Molecular , Xenopus/genética , beta-Galactosidase/genética , beta-Galactosidase/metabolismoRESUMO
Goosecoid is a homeobox gene that is expressed as an immediate early response to mesoderm induction by activin. We have investigated the induction of the zebrafish goosecoid promoter by the mesoderm inducing factors activin and basic fibroblast growth factor (bFGF) in dissociated zebrafish blastula cells, as well as by different wnts in intact embryos. Activin induces promoter activity, while bFGF shows a cooperative effect with activin. We have identified two enhancer elements that are functional in the induction of the goosecoid promoter. A distal element confers activin responsiveness to a heterologous promoter in the absence of de novo protein synthesis, whereas a proximal element responds only to a combination of activin and bFGF. Deletion experiments show that both elements are important for full induction by activin. Nuclear proteins that bind to these elements are expressed in blastula embryos, and competition experiments show that an octamer site in the activin responsive distal element is specifically bound, suggesting a role for an octamer binding factor in the regulation of goosecoid expression by activin. Experiments in intact embryos reveal that the proximal element contains sequences that respond to Xwnt1, but not to Xwnt5c. Furthermore, we show that the distal element is active in a confined dorsal domain in embryos and responds to overexpression of activin in vivo, as well as to dorsalization by lithium. The distal element is to our knowledge the first enhancer element identified that mediates the induction of a mesodermal gene by activin.
Assuntos
Proteínas de Ligação a DNA/genética , Fator 2 de Crescimento de Fibroblastos/fisiologia , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio , Inibinas/fisiologia , Regiões Promotoras Genéticas , Proteínas Repressoras , Fatores de Transcrição , Proteínas de Peixe-Zebra , Ativinas , Animais , Sequência de Bases , Blastocisto/citologia , Clonagem Molecular , Indução Embrionária , Elementos Facilitadores Genéticos , Proteína Goosecoid , Mesoderma/citologia , Dados de Sequência Molecular , Proteínas Nucleares/metabolismo , Proteínas/genética , Proteínas Proto-Oncogênicas/genética , Proteínas Wnt , Peixe-ZebraRESUMO
COUP-TFs (Chicken Ovalbumin Upstream Promoter Transcription Factors) have been proposed to be negative regulators of retinoid receptor-mediated transcriptional activation. In a previous paper we reported the cloning of a Xenopus (x) COUP-TF (Matharu, P.J. and Sweeney, G.E. (1992) Biochim. Biophys. Acta 1129, 331-334). Here we describe the cloning of a second xCOUP-TF. Amino acid sequence comparison between these two Xenopus COUP-TFs revealed a high level of similarity. Extensive amino acid sequence conservation was found among all Drosophila, Xenopus, zebrafish and mammalian COUP-TF genes examined. Phylogenetic tree analyses indicate that the vertebrate COUP-TFs fall into three classes. The two Xenopus COUP-TF genes show similar temporal expression patterns: both are expressed from the end of gastrulation. In situ hybridization studies reveal complex expression patterns in the developing central nervous system (CNS), besides expression in the eye and in some mesodermal tissues. Retinoic acid (RA) treatment enhances xCOUP-TF-A expression in neurula stage embryos, whereas the expression of xCOUP-TF-B is inhibited during the same developmental period. The strictly conserved amino acid sequences and the strong similarities between the expression patterns of the two different xCOUP-TFs on the one hand, and other vertebrate COUP-TF homologues on the other, make it likely that COUP-TFs have a conserved role in patterning the nervous system.
Assuntos
Proteínas de Ligação a DNA/genética , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Fatores de Transcrição/genética , Tretinoína/farmacologia , Sequência de Aminoácidos , Animais , Fator I de Transcrição COUP , Clonagem Molecular , DNA Complementar , Humanos , Dados de Sequência Molecular , Filogenia , Rombencéfalo/metabolismo , Especificidade da Espécie , Fatores de Tempo , Xenopus/genéticaRESUMO
As a first step towards the elucidation of the role of the transcription factor Oct-1 in development, we prepared a monoclonal antibody to study the spatio-temporal distribution of Oct-1 protein in vivo. Here we report differential expression of the Oct-1 gene in the Xenopus embryo both at the RNA and the protein level. Transcripts and protein are detected in ectodermal and mesodermal cell lineages, in which the expression exhibits a pattern of progressive spatial restriction in the course of development. The Oct-1 expression as reported here is not correlated with cell density or cell proliferation in the embryo. Our results suggest a role of Oct-1 in the specification and differentiation of neuronal and neural crest cells. In many other cells, the developmental decision to down regulate Oct-1 is delayed, probably due to a high stability of the protein.
Assuntos
Proteínas de Ligação a DNA/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , RNA/metabolismo , Fatores de Transcrição/metabolismo , Animais , Anticorpos Monoclonais , Divisão Celular , Linhagem Celular , Regulação para Baixo , Ectoderma/citologia , Embrião não Mamífero/metabolismo , Fator C1 de Célula Hospedeira , Fator 1 de Transcrição de Octâmero , Proteínas de Xenopus , Xenopus laevisRESUMO
The murine S8 gene, originally identified by Kongsuwan et al. [EMBO J. 7(1988)2131-2138] encodes a homeodomain which resembles those of the paired family. We studied the expression pattern during mid-gestation embryogenesis of S8 by in situ hybridization. Expression was detected locally in craniofacial mesenchyme, in the limb, the heart and the somites and sclerotomes all along the axis, and was absent from the central and peripheral nervous system, splanchnopleure, and endodermal derivatives. This pattern differs considerably from that of most previously described homeobox containing genes. By genetic analysis, the gene was located on chromosome 2, about 20 cM from the HOX-4 cluster.
Assuntos
Genes Homeobox/fisiologia , Mesoderma/metabolismo , Camundongos/embriologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Northern Blotting , Extremidades/embriologia , Expressão Gênica , Cabeça/embriologia , Dados de Sequência Molecular , Miocárdio/metabolismo , Hibridização de Ácido Nucleico , Sondas RNA , Mapeamento por Restrição , Homologia de Sequência do Ácido NucleicoRESUMO
Here, we review the WNT pathway and its regulation at different levels. We focus on the transcriptional regulation of WNT target genes, in light of the recently identified negative regulators, i.e. relatives of groucho and CBP.
Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Proteínas Proto-Oncogênicas/fisiologia , Transativadores , Proteínas de Peixe-Zebra , Animais , Proteínas do Citoesqueleto/metabolismo , Proteínas de Ligação a DNA/fisiologia , Embrião não Mamífero , Fator 1 de Ligação ao Facilitador Linfoide , Proteínas Proto-Oncogênicas/metabolismo , Fatores de Transcrição/fisiologia , Proteínas Wnt , Xenopus , Proteínas de Xenopus , beta CateninaRESUMO
Expression of the Engrailed-1 (XEn-1) gene was studied in Xenopus embryogenesis by Northern blot analysis and whole-mount in situ hybridization. One transcript of 2.2 kb was detected from stage 17 (midneurula) onwards, until stage 47 (swimming tadpole). The expression pattern of the XEn-1 gene as revealed by in situ hybridization can be divided in three regions. The first domain of transient expression appears at the midneurula stage (st. 17) in the anterior part of the neural fold, forming a complete ring of positive cells at the mid/hindbrain border after neural tube closure. A second region of transient expression is detected as groups of ventro-lateral cells in the spinal cord and the hindbrain from late-neurula till tadpole stages. A third area of transient expression of XEn-1 is formed by the anterior part of the developing pronephros. Comparison of XEn-1 expression at the mid/hindbrain border with that of the Xenopus wnt-1 and engrailed-2 genes reveals that XEn-1 and Xwnt-1, in contrast to XEn-2, are both detected in a narrow stripe of positive cells in this region. Analysis in exogastrulated embryos reveals that expression of XEn-1 and Xwnt-1, but not XEn-2, is induced by planar signaling in the presumptive midbrain. Of the three genes only XEn-1 is expressed in the floorplate at the mid/hindbrain border, while Xwnt-1 is expressed in adjacent cells in the neural ectoderm. The results suggest that in vertebrates at the interface between cells in the floorplate and in the paraxial neuroectoderm, at the limited region of the mid/hindbrain border, En-1 interacts with wnt-1 in a signaling pathway analogous to the engrailed/wingless signaling in the parasegments of the Drosophila embryo.