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Title: Characterisation of a novel gene (Dstpk61) encoding a serine/threonine protein kinase in Drosophila melanogaster
Author: Clyde, Dorothy
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 1999
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Only a few targets of the sex determining genes are known - dsf, fru, and the yps. Such genes would be expected to fit into one of three categories: (i) Genes expressed exclusively in one sex; (ii) Genes expressed at a higher level in one sex than the other; (iii) Genes that are expressed at similar levels in both sexes, but produce sex-specific transcripts. In an attempt to isolate genes that fit into these categories a non-gonadal differential screen of an adult body genomic DNA library was performed, using sex-specific radio-labelled cDNA. The genomic recombinant fs(1) was chosen for further analysis based on its preferential hybridisation to female non-gonadal cDNA. Screening of cDNA libraries using fs(1) sequence isolated two cDNAs, a 3.0kb cDNA (cDNAA) and a 4.5kb cDNA (cDNAB). Sequence analysis revealed that both these cDNAs encoded a novel opa repeat-containing serine/threonine protein kinase, the difference in the cDNAs being the length of the 5' and 3' untranslated regions (UTRs). The gene, which maps to polytene position 61A, was named Drosophila serine/threonine protein kinase@61 (Dstpk61). Northern analysis showed there to be at least 4 different Dstpk61 transcripts: a 3.0kb testis-specific transcript (possibly represented by cDNAA); a 3.5kb ovary-specific transcript; a 4.5kb female-specific carcass transcript (possibly represented by cDNAB); and a slightly larger non-sex-specific transcript. The work presented here shows that Dstpk61 is a complex gene that produces multiple transcripts by using alternative promoters, alternative polyadenylation sites, and alternative splicing. It is now known to have significant sequence and functional homology to mammalian Phosphoinositol-dependent kinase 1 (PDK1), which has been implicated in the insulin signal transduction pathway and the regulation of apoptosis. Four mutant P-insertion lines have been identified that disrupt Dstpk61 at different sites. The phenotypes of these mutants, possible Dstpk61 transcripts, and the observed mRNA expression pattern are discussed in relation to the possible role of Dstpk61 protein in Drosophila development.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available