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Identification of hox-regulated targets at different time points of a developmental process


Hox genes are present in all animals with bilateral symmetry, providing a conserved mechanism to control regional differences along the body axis. However, little is known about the sets of genes that are modulated by the Hox proteins to mould morphologically distinct structures. The aim of this proposal is to analyse how the Hox proteins impinge on regulatory networks executing cell differentiation programmes. To study such a complex biological process, I will use as an experimental system the homeotic transformation of wing cells to haltere cells by the Hox protein Ultrabithorax (Ubx) in Drosophila melanogaster. By means of hybridisations to DNA microarrays, I will compare the transcriptome of control and Ubx-expressing wing discs. The sensitivity of these experiments will be enhanced by collecting additional microarray datasets from wing and haltere discs expressing an engineered form of Ubx with a potent transcriptional activation function. The way cells respond to Ubx changes several times during imaginal disc development. By employing a sophisticated expression system in Drosophila, I will identify the transcriptional targets of Ubx at different time points during larval and pupal development. All experiments will be done in duplicates, both with in vivo -processed discs and with discs cultured in vitro in the presence of the protein synthesis inhibitor cycloheximide. This will allow to distinguish between direct and indirect targets of Ubx. The biological significance of candidate targets will be verified by molecular genetic techniques and the relevant cis-regulatory regions will be retrieved by computational approaches. I will make use of this project to receive advanced training in complex genetic manipulations, whole-genome expression profiling, comparative genomics and bioinformatics. This research activity will be the foundation stone in a long-term work plan that will assist my career advancement in the field of Evolutionary Developmental Biology.

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The Old Schools, Trinity Lane
United Kingdom

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