Analysis of Rap1 on protrusion dynamics in Drosophila S2 Cell

Abstract

Rap1 belongs to the Ras superfamily of small GTPase which is essential for cell migration and the embryonic development Previous study in our lab found that the Rap1 was required for border cell migration especially in regulation of asymmetrical distributions of actin at the leading edge of the migration cluster To further examine the general mechanism of Rap1 in actin dynamics we analyzed the cellular protrusions in Drosophila S2 cells by transfections of the Rap1 Rap1V12 or Rap1N17 Interestingly we observed 71% of S2 cells in overexpression of Rap1V12 showing Moreover we also observed long protrusions that extended to 18 8 μm in length in expression of Rap1V12 In time lapse micrographs we found that the Rap1V12 transfected S2 Mt-Actin::GFP cells could induce extended long protrusions and multiple protrusions which was consistent with the results observed in fix samples In our speed analysis we found that the Rap1V12 transfected cell had more high displayed faster speed no matter in protrusion withdraw or extension We further investigate several actin binding proteins to explore the role of Rap1 in actin dynamics We found that the expression of WASP and SCAR was higher in Rap1V12 transfected cells Taken together our finding suggests that active form of Rap1 might regulate protrusion dynamics through SCAR and WASP

Date of Award	2014 Sept 9
Original language	English
Supervisor	Chuen-Chuen Jang (Supervisor)