RESUMEN
Runx1 (also known as AML1, Cbfa2 and Pebpa2b) and Cbfb encode a DNA-binding alpha subunit and the non-DNA-binding beta subunit of a mammalian core-binding factor (CBF). The discovery of RUNX1 and CBFB as genes rearranged in human leukemias prompted predictions that both genes would play important roles in normal hematopoiesis. These predictions were borne out, as indeed Runx1 and its Xenopus and Drosophila homologs, Xaml and lozenge (lz), appear to determine hematopoietic cell fate during development. We will review what is known about Runx1 function in hematopoiesis in three model organisms, mouse, frog and fly, focusing on the earliest events of hematopoietic cell emergence in the embryo.
Asunto(s)
Proteínas de Unión al ADN/metabolismo , Proteínas de Drosophila , Hematopoyesis , Proteínas Proto-Oncogénicas , Factores de Transcripción/metabolismo , Proteínas de Xenopus , Animales , Subunidad alfa 2 del Factor de Unión al Sitio Principal , Proteínas de Unión al ADN/genética , Drosophila melanogaster/embriología , Drosophila melanogaster/genética , Drosophila melanogaster/crecimiento & desarrollo , Desarrollo Embrionario y Fetal/genética , Desarrollo Embrionario y Fetal/fisiología , Hematopoyesis/genética , Humanos , Factores de Transcripción/genética , Xenopus laevis/embriología , Xenopus laevis/genética , Xenopus laevis/crecimiento & desarrolloRESUMEN
The specific functions of gene products frequently depend on the developmental context in which they are expressed. Thus, studies on gene function will benefit from systems that allow for manipulation of gene expression within model systems where the developmental context is well defined. Here we describe a system that allows for genetically controlled overexpression of any gene of interest under normal physiological conditions in the early Drosophila embryo. This regulated expression is achieved through the use of Drosophila lines that express a maternal mRNA for the yeast transcription factor GAL4. Embryos derived from females that express GAL4 maternally activate GAL4-dependent UAS transgenes at uniform levels throughout the embryo during the blastoderm stage of embryogenesis. The expression levels can be quantitatively manipulated through the use of lines that have different levels of maternal GAL4 activity. Specific phenotypes are produced by expression of a number of different developmental regulators with this system, including genes that normally do not function during Drosophila embryogenesis. Analysis of the response to overexpression of runt provides evidence that this pair-rule segmentation gene has a direct role in repressing transcription of the segment-polarity gene engrailed. The maternal GAL4 system will have applications both for the measurement of gene activity in reverse genetic experiments as well as for the identification of genetic factors that have quantitative effects on gene function in vivo.
Asunto(s)
Drosophila/genética , Embrión no Mamífero , Proteínas de Saccharomyces cerevisiae , Animales , Proteínas de Unión al ADN , Drosophila/embriología , Proteínas Fúngicas/genética , Regulación de la Expresión Génica , Vectores Genéticos , Impresión Genómica , Fenotipo , Regiones Promotoras Genéticas , ARN Mensajero/genética , Factores de Transcripción/genética , TransgenesRESUMEN
The Runt domain gene AML1 is essential for definitive hematopoiesis during murine embryogenesis. We have isolated Xaml, a Xenopus AML1 homologue in order to investigate the patterning mechanisms responsible for the generation of hematopoietic precursors. Xaml is expressed early in the developing ventral blood island in a pattern that anticipates that of later globin. Analysis of globin and Xaml expression in explants, in embryos with perturbed dorsal ventral patterning, and by lineage tracing indicates that the formation of the ventral blood island is more complex than previously thought and involves contributions from both dorsal and ventral tissues. A truncated Xaml protein interferes with primitive hematopoiesis. Based on these results, we propose that Runt domain proteins function in the specification of hematopoietic stem cells in vertebrate embryos.