Genetics - Theses

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Author: Smith, Michiko. × End date: 1999 × Start date: 1980 ×

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    Molecular and cellular biological studies of the Drosophila melanogaster stoned gene
    Smith, Michiko. (University of Melbourne, 1999)
    The Drosophila melanogaster stoned locus is an essential neurologically expressed gene, of which temperature-sensitive mutants exhibit a lethal interaction with the shibire locus. This, and subsequent evidence gathered from electrophysiological, localisation and sequence analyses of stoned have indicated a likely role critical to the synaptic vesicle cycle. Taking advantage of the genetic tools available for the dissection of stoned function, the role of stoned in the synaptic vesicle cycle was directly implicated in this study. The stoned transcript has a unique tandemly-arranged dicistronic structure, with both open reading frames (ORFs) separated by a fifty-five nucleotide sequence. Presented in this study, is firstly the identification of two distinct translatory products of the stoned ORFs, StonedA and StonedB. Although both novel proteins, the predicted amino acid sequences have allowed for the progressive identification of domains that have homology to a number of proteins implicated in the synaptic vesicle cycle. StonedA and StonedB were determined to have a direct association with synaptic vesicles by co-immunoprecipitation and sedimentation properties. However, rather than a ubiquitous association with synaptic vesicles, StonedA and StonedB are associated with separate subsets within the entire population. The biochemical localisation of StonedB repartitioned over D. melanogaster shibire background subcellular fractions. The data suggests that StonedA and StonedB are indeed part of the molecular mechanisms in the synaptic vesicle cycle. Perhaps StonedB-tagged synaptic vesicles are involved in the elusive kiss-and-run mechanism of the synaptic vesicle cycle.
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    Molecular and cellular biological studies of the Drosophila melanogaster stoned gene
    Smith, Michiko. (University of Melbourne, 1999)
    The Drosophila melanogaster stoned locus is an essential neurologically expressed gene, of which temperature-sensitive mutants exhibit a lethal interaction with the shibire locus. This, and subsequent evidence gathered from electrophysiological, localisation and sequence analyses of stoned have indicated a likely role critical to the synaptic vesicle cycle. Taking advantage of the genetic tools available for the dissection of stoned function, the role of stoned in the synaptic vesicle cycle was directly implicated in this study. The stoned transcript has a unique tandemly-arranged dicistronic structure, with both open reading frames (ORFs) separated by a fifty-five nucleotide sequence. Presented in this study, is firstly the identification of two distinct translatory products of the stoned ORFs, StonedA and StonedB. Although both novel proteins, the predicted amino acid sequences have allowed for the progressive identification of domains that have homology to a number of proteins implicated in the synaptic vesicle cycle. StonedA and StonedB were determined to have a direct association with synaptic vesicles by co-immunoprecipitation and sedimentation properties. However, rather than a ubiquitous association with synaptic vesicles, StonedA and StonedB are associated with separate subsets within the entire population. The biochemical localisation of StonedB repartitioned over D. melanogaster shibire background subcellular fractions. The data suggests that StonedA and StonedB are indeed part of the molecular mechanisms in the synaptic vesicle cycle. Perhaps StonedB-tagged synaptic vesicles are involved in the elusive kiss-and-run mechanism of the synaptic vesicle cycle.