RESUMEN
In eye development the tasks of tissue specification and cell proliferation are regulated, in part, by the Pax6 and Pax6(5a) proteins respectively. In vertebrates, Pax6(5a) is generated as an alternately spliced isoform of Pax6. This stands in contrast to the fruit fly, Drosophila melanogaster, which has two Pax6(5a) homologs that are encoded by the eyegone and twin of eyegone genes. In this report we set out to determine the respective contributions that each gene makes to the development of the fly retina. Here we demonstrate that both eyg and toe encode transcriptional repressors, are expressed in identical patterns but at significantly different levels. We further show, through a molecular dissection of both proteins, that Eyg makes differential use of several domains when compared to Toe and that the number of repressor domains also differs between the two Pax6(5a) homologs. We predict that these results will have implications for elucidating the functional differences between closely related members of other Pax subclasses.
Asunto(s)
Proteínas de Unión al ADN/genética , Proteínas de Drosophila/genética , Drosophila melanogaster/crecimiento & desarrollo , Drosophila melanogaster/genética , Proteínas del Ojo/genética , Ojo/crecimiento & desarrollo , Genes de Insecto , Proteínas de Homeodominio/genética , Factor de Transcripción PAX5/genética , Factores de Transcripción Paired Box/genética , Proteínas Represoras/genética , Animales , Animales Modificados Genéticamente , Secuencia de Bases , ADN/genética , ADN/metabolismo , Cartilla de ADN/genética , Proteínas de Unión al ADN/química , Proteínas de Drosophila/química , Drosophila melanogaster/embriología , Drosophila melanogaster/metabolismo , Ojo/embriología , Ojo/metabolismo , Proteínas del Ojo/química , Regulación del Desarrollo de la Expresión Génica , Proteínas de Homeodominio/química , Hibridación in Situ , Factor de Transcripción PAX5/química , Factor de Transcripción PAX6 , Factores de Transcripción Paired Box/química , Estructura Terciaria de Proteína , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Recombinantes de Fusión/genética , Proteínas Represoras/químicaRESUMEN
The initiation of retinal development in Drosophila begins at the posterior center (PC) of the eye disc margin. The front of the differentiation wave, recognized as a morphogenetic furrow (MF), moves from posterior to anterior. What determines MF initiates from the specific PC site is still unclear. The unpaired (upd) gene is expressed at PC at early third instar, just before the time of MF initiation. Therefore, upd is expressed at the appropriate time and location for a specific role in defining the site of MF initiation. upd encodes a ligand for the Jak/STAT signaling pathway. In this report, we showed that the Upd/Jak/STAT signaling is required and sufficient to determine MF initiation. This is primarily achieved by repressing the transcription of wingless (wg), which is known to block MF initiation.
Asunto(s)
Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Regulación del Desarrollo de la Expresión Génica , Janus Quinasa 1/metabolismo , Células Fotorreceptoras de Invertebrados/embriología , Proteínas Proto-Oncogénicas/genética , Transducción de Señal , Factores de Transcripción/metabolismo , Transcripción Genética , Animales , Animales Modificados Genéticamente , Diferenciación Celular , Drosophila melanogaster/embriología , Inmunohistoquímica , Ligandos , Células Fotorreceptoras de Invertebrados/metabolismo , Proteínas Proto-Oncogénicas/metabolismo , Factores de Transcripción STAT/metabolismo , Proteína Wnt1RESUMEN
Drosophila has two pairs of Pax genes, ey/toy and eyg/toe, that play different functions during eye development. ey specifies eye fate, while eyg promotes cell proliferation. We have determined the molecular basis for the functional diversity of Eyg and Ey. Eyg and Ey act by distinct transcriptional mechanisms. They use different DNA-binding domains for target recognition. Most interestingly, Eyg acts exclusively as a repressor, whereas Ey is an activator. Several vertebrate Pax proteins are known to switch between activator and repressor activities, but none as repressors only. Eyg may be the first Pax protein as a dedicated repressor. Vertebrates produce a Pax6 isoform, Pax6-5a, differing from Pax6 in DNA-binding properties and functions and structurally similar to Eyg/Toe. We found that Pax6-5a acts as an activator like Ey, but has DNA-binding specificity like Eyg.