TUPLE1 is a putative transcriptional factor which has been isolated from the DiGeorge critical region. Haploinsuffiency for TUPLE1 could be partly responsible for DGS and related abnormalities. TUPLE1 is highly conserved across many species including mammals, tish and insects.
The work I propose is to isolate TUPLE1 homologues from Fugu (puffer fish) and Drosophila. This approach will allow the identification of conserved or functional motifs. The human TUPLE1 cDNA has been isolated and sequenced. The complete gene structure and flanking regions of the Fugu TUPLE1 homologue will be determined by sequencing from genomic DNA then the sequence will be compared with the human gene. A similar analysis will be done in Drosophila. Conserved elements between the species will provide clues to the functional relevance of particular motifs and provide a basis on which to begin experiments to determine function. Comparisons among species may identify important conserved features in the promoter and upstream flanking region of TUPLE1 which could be the first candidates for mutagenesis studies using Drosophila as a model system.
In addition, with the isolation of TUPLE1 from the fish and fly we can compare the evolution of TUPLE1 in human, mouse, chick, fish, frog, fly and yeast.