Systems Analysis of G-protein dynamics in D. discoideum; a pilot study using novel 3D microscopy computational modelling and micromanipulation

Lead Research Organisation: University of Edinburgh
Department Name: Sch of Biological Sciences


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Description 1. A pilot study was performed in the group of Ass. Prof. C. Janetopoulos, VU. The main directions of the experimental work included: TIRF FRAP of G protein subunits in live D. d. cells to determine their trafficking parameters on the membrane and cytoplasm; design of a reagent for the detection of activated G-alpha subunits based on G-alpha effector proteins; design of constitutively active G-alpha subunits; generation of G- betta/gamma subunits fused to the FKBP-rapamycine interacting components for rapid sequestration of G-betta/gamma subunits; MS proteomics approach to identify interactome of G- protein subunits.
2. The group of Dr. A. Goryachev, U of E, performed extensive data analysis of the data generated by the VU collaborators, in particular, the results of the FRAP experiments. A prototype model of G-protein trafficking has been initiated based on the experimental results and will be completed pending the results of ongoing experiments.
3. The intermediate results of the project were disseminated at the organized by us 3-day workshop that took place on the premises of VU and brought together prominent researchers in cell migration, live cell microscopy and imaging from the US and UK. The total number of participants reached 90 people. The detailed programme of the meeting including the titles of all talks is available at
4. An ongoing dialogue between the groups of Drs. C. Janetopoulos and A. Goryachev, the basis for long term collaboration, has been firmly established. Further work and communication based on the pilot project supported by BBSRC is planned. Pending publication of the early results, joint funding application will be submitted in the US and UK.
Exploitation Route This was a short term partnering award intended to support travel and communication.
Sectors Education,Healthcare,Pharmaceuticals and Medical Biotechnology

Description Collaboration with Prof. C. Janetopoulos 
Organisation Vanderbilt University
Department Department of Cell and Developmental Biology
Country United States 
Sector Academic/University 
PI Contribution Data analysis and mathematical modeling
Collaborator Contribution Experimental data in Dictyostelium amoeba
Impact The main publically-visible output of this collaborative project was the organized by us three day workshop "Cell signaling and cytoskeleton in directed cell migration: Imaging and quantitative approaches" that brought together experimentalists and computational modelers from the US and UK. The meeting took place in VU, Marc 4-6, 2012 and attracted close to 90 participants. The detailed programme of the meeting including the titles of all talks is available at UK science was represented by Profs. Kurt Anderson, Robert Insall (Beatson Cancer Research Institute, Glasgow) and Drs. Angeliki Malliri (Patterson Cancer Research Institute, Manchester) and A. Goryachev (U of Edinburgh). Three days of invited and contributed talks were exceptionally well attended. High interest in this meeting was reflected in a generous contribution from the Vanderbilt University departments which amounted to $5,500.00.
Start Year 2009