Analysis and reconstitution of the genetic cascade controlling early mesoderm morphogenesis in the Drosophila embryo.

To understand how transcription factors direct developmental events, it is necessary to know their target or 'effector' genes whose products mediate the downstream cell biological events. Whereas loss of a single target may partially or fully recapitulate the phenotype of loss of the transcription factor, this does not mean that this target is the only direct mediator. For a complete understanding of the pathway it is necessary to identify the full set of targets that together are sufficient to carry out the programme initiated by the transcription factor, which has not yet been attempted for any pathway. In the case of the transcriptional activator Twist, which acts at the top of the mesodermal developmental cascade in Drosophila, two targets, Snail and Fog, are known to be necessary for the first morphogenetic event, the orderly invagination of the mesoderm. We use a system of reconstituting loss of Twist function by transgenes expressing Snail and Fog independently of Twist to analyse the sufficiency of these factors-a loss of function assay for additional gene functions to assess what further functions might be needed downstream of Twist. Confirming and extending previous studies, we show that Snail plays an essential role, allowing basic cell shape changes to take place. Fog and at least two other genes are needed to accelerate and coordinate shape changes. Furthermore, this study represents the first step in the systematic reconstruction of the morphogenetic programme downstream of Twist.