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Title: Drosophila homologs of HIV-1 Rev interacting proteins
Author: Chan, H. E.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 1999
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The aim of this project was to analyze four RIPs (Crml, Drongo, Dribble and eIF5A) in Drosophila melanogaster. Two RIPs, Drongo and Dribble, are encoded by genes that lie about 20 kb apart in the 21D/E region of the Drosophila genome. I have generated a transcript map of this region, identified the locations of several P insertions, and mapped a number of mutations in the region. I have generated null mutations in one RIP gene, dribble (dbe). Dribble is the Drosophila homologue of Human Rev-interacting protein 1. The Drosophila dbe gene encodes a novel KH domain protein. Maternal expression of dbe transcripts was detected during oogenesis while zygotic dbe expression is ubiquitous at low levels throughout embryogenesis. Dribble protein was detected essentially in the cell nucleus. All existing dbe mutants die at first instar larval stage and display no obvious phenotype. In a homozygous dbe mutant background, both a Rev-GFP and a nuclear localization signal peptide-GFP fusion protein were retained in the nucleus. Therefore Dribble is not required for the nuclear import or retention of these proteins. To study the effect of dbe on protein nuclear export, I developed a novel nuclear export assay in Drosophila. In this assay, the subcellular localization of endogenous cytoplasmic actin is used as an export indicator. A putative nuclear export mutant crmI was used as a positive control. As expected, nuclear accumulation of actin was observed in crmI mutants. This finding showed that crmI is required for nuclear export in Drosophila. In a homozygous dbe mutant background, wild-type cytoplasmic localization of actin was observed, suggesting that dbe is not essential for protein nuclear export. I have identified and mapped the Drosophila homolog of the eIF5A gene in the 60B genomic region, and identified a P insertion in the first intron of the gene.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available