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Title: Genetic dissection of meiotic cytokinesis in Drosophila melanogaster males
Tutor: Giansanti, Maria Grazia
Keywords: cytokinesis
central spindle
contractile ring
Issue Date: 25-Feb-2013
Abstract: Studies in a variety of organisms indicate that membrane traffic to the cleavage furrow is an essential facet of cytokinesis involving components of the secretory and endocytic/recycling trafficking pathways, as well as the membrane fusion machinery. My PhD project focused around the characterization of two genes required for cleavage furrow formation and ingression in male meiotic cells namely sauron and Cog7 which encode the Drosophila orthologues of two known players of membrane trafficking pathways. Sauron is the Drosophila orthologue of Golgi phosphoprotein 3 (GOLPH3) which has recently been recognized as a potent oncogene amplified in many human cancers. Studies in both yeast and mammalian cells have implicated this protein in several vesicle trafficking events. I show that Sau/dGOLPH3 is required during the early steps of spermatocyte cytokinesis playing an essential role in both contractile ring assembly and vesicle trafficking during furrow ingression. Consistently my work suggests that Sau/dGOLPH3 might interact with components of both the contractile apparatus and the membrane trafficking machinery in male germ cells. The conserved oligomeric Golgi (COG) Complex plays essential roles for Golgi function, vesicle trafficking and glycosylation. Mutations in the genes encoding human COG1, COG4-COG8 have been associated with congenital disorders of glycosylation (CDG). Deletions of human COG7 are associated with a rare multisystemic congenital disorder of glycosylation causing mortality within the first year of life. I show that, similar to Drosophila Cog5, Cog7 controls furrow ingression during cytokinesis. Importantly, Cog7 is required to localize the small GTPase Rab11 and the phosphatidylinositol transfer protein (PITP) Giotto (Gio) to the cleavage site of spermatocytes. In addition Gio coimmunoprecipitates with both Cog7 and Rab11 in Drosophila testes suggesting that these proteins may interact in male germ cells.
Research interests: Dr Sechi’s research is aimed at elucidating the molecular mechanism coupling membrane remodelling and cytoskeletal dynamics during cytokinesis using Drosophila melanogaster as a model system. Specifically, Dr. Sechi is interested in understanding the mechanisms underlying the formation of actomyosin ring and its interaction with the microtubules of the central spindle during the early stages of cytokinesis, processes which shed light on the role of membrane trafficking and remodeling during cleavage furrow ingression
Skills short description: Dr. Sechi has expertise in formal genetics of Drosophila melanogaster, including proficiency in genetic mapping, P-element mobilization to induce genetic mutations, and studies of genetic interactions. In the field of microscopy, Dr. Sechi has skills related to the analysis of Drosophila tissue (spermotocytes and neuroblasts) and cultured cells (S2 cells) of Drosophila melanogaster fixed for indirect immunofluorescence, as well as in vivo assays using time-lapse videomicroscopy. In the field of molecular biology Dr. Sechi has expertise with protocols utilizing gene cloning technology as well as expression of recombinant proteins. Furthermore, Dr. Sechi is also capable of studying proteins on a molecular level, using co-immunoprecipitation and GST-Pull Down to assay intermolecular interactions and multiple bioinformatics approaches for the comparison of amino acid sequences and structure-function prediction.
Personal skills keywords: Drosophila Genetics
Gene cloning
Analysis of protein interaction (GST-pull down and Co-immunoprecipitation experiments)
Time lapse-video microscopy using Metamorph software and a confocal spinning disk
Bioinformatics expertise

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