RESUMEN
Bid protein, a member of the "BH3-only" subgroup of Bcl-2 family, plays a critical role in mammalian apoptosis regulation. In this study, we have cloned the chicken Bid gene, which encodes a 193 amino acid protein and shares 40% homology with human and mouse Bid proteins. Bid sequence comparison emphasises the conservation of both the functional domain BH3 and the proteolytic cleavage sites. An induction of apoptosis by chicken Bid and the cleavage of the protein, after TNFalpha treatment, were also demonstrated. In addition, mRNA Bid expression was detected along all embryo stages and tissues examined, suggesting a role for this protein in the developmental process. This is the first report demonstrating the functionality of a "BH3-only" protein in chicken.
Asunto(s)
Proteína Proapoptótica que Interacciona Mediante Dominios BH3/metabolismo , Secuencia de Aminoácidos , Animales , Apoptosis/efectos de los fármacos , Proteína Proapoptótica que Interacciona Mediante Dominios BH3/química , Proteína Proapoptótica que Interacciona Mediante Dominios BH3/genética , Embrión de Pollo , Pollos , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Humanos , Datos de Secuencia Molecular , Filogenia , Procesamiento Proteico-Postraduccional , ARN Mensajero/genética , ARN Mensajero/metabolismo , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
The chicken (Gallus gallus) is one of the primary models for embryological and developmental studies. In order to begin to understand the molecular mechanisms underlying the normal and abnormal development of the chicken, we used 2-DE to construct a whole-embryo proteome map. Proteins were separated by IEF on IPG strips, and by 11% SDS-PAGE) gels. Protein identification was performed by means of PMF with MALDI-TOF-MS. In all, 105 protein spots were identified, 35 of them implicated in embryo development, 10 related with some diseases, and 16, finally, being proteins that have never been identified, purified or characterized in the chicken before. This map will be updated continuously and will serve as a reference database for investigators, studying changes at the protein level under different physiological conditions.
Asunto(s)
Embrión de Pollo/crecimiento & desarrollo , Proteoma/análisis , Animales , Embrión de Pollo/metabolismo , Electroforesis en Gel Bidimensional/métodos , Regulación del Desarrollo de la Expresión Génica/fisiologíaRESUMEN
Nup88 is a nuclear pore complex protein which is overexpressed in a variety of human tumors of the stomach, colon, liver, pancreas, breast, lung, ovary, uterus, prostate and kidney. A monoclonal antibody crossreacting with the yeast Candida albicans and Nup88 was used to investigate the expression of cross-reactive antigens in chick embryos, in an attempt to identify an experimental model for studying the role played by Nup88 during cell development and differentiation. All cells in the trilaminar embryo were labeled with the antibody, but as development advanced and organogenesis was completed, expression of the corresponding antigen became more restricted. Thus, some structures continued to be intensely labeled (skin epithelium, oropharyngeal endothelium, perichondral mesenchymal tissue), whereas others ( muscular tissue, vascular endothelium, respiratory endothelium, digestive tract mucosa, peripheral nerves, medullary white matter and the retinal axons) were more moderately stained. No immunoreactivity was observed in the medullary grey matter or cartilage. A specific band of 53 kDa observed by Western blotting of chick embryo extracts suggested that the chicken antigen recognized by the monoclonal antibody is the homologue of human Nup88, which is associated with the high proliferation and low differentiation of tumor cells. The present results indicate that the role of Nup88 in cell differentiation and organ development could be fruitfully investigated using the developing chick embryo as an experimental model.
