Dahlsveen, Ina Kathrine; (2002) Identification and characterisation of proteins that interact with the Drosophila transcription factor mirror. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

Mirror is a homeodomain transcription factor in Drosophila melanogaster. The mirror gene is part of the Iroquois complex (Iro-C) which also contains araucan and caupolican. The Iro-C genes have important roles in the development and patterning of the eye and wing imaginal discs as well as the oocyte and embryo. The Iroquois gene products are the founding members of the IRO family of proteins which are characterised by a highly conserved homeodomain of the TALE class (three amino acid loop extension). IRO proteins have been identified in organisms from C. elegans to humans and seem to have conserved roles in developmental patterning. To discover more about how Mirror functions as a transcription factor, the yeast-two-hybrid system was used to screen a Drosophila embryonic cDNA library for proteins that interact with Mirror. Several putative interactors were identified including two known transcription factors and a chromatin-remodelling factor as well as a number of previously uncharacterised gene products. A subset of the confirmed yeast-two-hybrid interactors have been investigated for biological relevance by comparing their expression patterns and mutant phenotypes to those of mirror as well as through genetic interactions. Two of the putative Mirror-binding proteins have been studied in detail, the novel forkhead associated (FHA) domain containing protein CG1135 and chromodomain helicase/ATPase DNA-binding protein 1 (CHD1). A P element insertion allele of CG1135 was shown to interact genetically with mirror alleles. Characterisation of the CG1135 phenotype revealed a putative role in cell proliferation or survival. The function of CHD1 in Drosophila is not known. In order to characterise the role of CHD1 and investigate any interactions with Mirror, a putative dominant negative version was generated and analysed. Studies of the localisation of CHD1 on polytene chromosomes indicate that CHD1 may be associated with transcriptional elongation.

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