Final Report Summary - DASPP/BOA PROTEINS (Functional Analysis of dASPP/Boa, novel regulators of dCsk in epithelial growth and morphogenesis)
Final Publishable Summary Report
Please see attached document for porject objectives and work performed.
A description of the main results achieved so far
We previously identified boa/dASPP complex as novel AJ component required for the proper localisation of DE-cadherin. I set out to elucidate the mechanism by which Boa affects AJ stability and found that Boa binds to Sec15, an exocyst component regulating polarised vesicle trafficking. Similar to Sec15 mutants, Boa mutants show an accumulation of Rab11-containing recycling endosomes, suggesting defects in the vesicle trafficking pathway. Boa also genetically interacts with Sec15. Electron microscopy data shows that Boa mutants seem to have a defect in the organization of the vesicle sorting pathway. In order to study the dynamics of the AJs and vesicles in living animal, we have now set up live-imaging and FRAP analysis in developing mosaic pupal retinas which allows us to perform clonal analysis live.
Expected final results
Once I finish performing the in vivo live-imaging/ FRAP experiments and analysing the data, I will have quantitative data to show the effect of Boa in AJs and vesicle dynamics. The results from the 3D TEM experiment will provide further information on the defect of vesicle trafficking pathway in Boa mutants. I will then have a complete picture of the functions of Boa/sec15 complex in AJs maintenance. My findings show that Boa, together with Sec15 modulates polarised vesicle trafficking and thus AJs stability during epithelial morphogenesis.
Please see attached document for porject objectives and work performed.
A description of the main results achieved so far
We previously identified boa/dASPP complex as novel AJ component required for the proper localisation of DE-cadherin. I set out to elucidate the mechanism by which Boa affects AJ stability and found that Boa binds to Sec15, an exocyst component regulating polarised vesicle trafficking. Similar to Sec15 mutants, Boa mutants show an accumulation of Rab11-containing recycling endosomes, suggesting defects in the vesicle trafficking pathway. Boa also genetically interacts with Sec15. Electron microscopy data shows that Boa mutants seem to have a defect in the organization of the vesicle sorting pathway. In order to study the dynamics of the AJs and vesicles in living animal, we have now set up live-imaging and FRAP analysis in developing mosaic pupal retinas which allows us to perform clonal analysis live.
Expected final results
Once I finish performing the in vivo live-imaging/ FRAP experiments and analysing the data, I will have quantitative data to show the effect of Boa in AJs and vesicle dynamics. The results from the 3D TEM experiment will provide further information on the defect of vesicle trafficking pathway in Boa mutants. I will then have a complete picture of the functions of Boa/sec15 complex in AJs maintenance. My findings show that Boa, together with Sec15 modulates polarised vesicle trafficking and thus AJs stability during epithelial morphogenesis.