Project properties
Title The role of the actin binding protein Actin Depolymerising Factor (ADF) in actin organisation, actin dynamics and plant development analysed by inducible inhibition of ADF
Group Plant Cell Biology, Laboratory of
Project type thesis
Credits 18-39
Supervisor(s) Dr. Tijs Ketelaar
Examiner(s) Prof. Dr. Marcel Janson
Contact info tijs.ketelaar@wur.nl Phone: 0317 482453
Begin date 2008/04/01
End date 2010/04/01
Description The actin cytoskeleton is involved in organisation of the cytoplasm, intracellular transport and regulation of cell growth. It is a dynamic structure that changes its organisation by remodelling, growing and shrinking constantly. The dynamics of the actin cytoskeleton are controlled by many actin associated proteins. One of these proteins is Actin Depolymerising Factor (ADF). ADF binds to actin filaments, cuts them into smaller pieces and promotes depolymerisation of the severed fragments in a pH dependent manner. The knowledge we have about ADF comes mainly from in vitro experiments. It is not known to what extent ADF contributes to the dynamics of the actin cytoskeleton in plants and what consequences the inhibition of ADF has on actin dynamics and actin organisation. To understand the consequences of the regulation of the actin cytoskeleton in live plants by ADF, we will inhibit ADF expression in Arabidopsis plants using an ethanol inducible RNA interference (RNAi) approach. The actin cytoskeleton will be visualised by expressing a fusion of Green Fluorescent Protein (GFP) with an actin binding domain (GFP-ABD). The levels of ADF in GFP-ABD expressing Arabidopsis plants, transformed with the ethanol inducible ADF RNAi construct, will be reduced by induction of the ADF RNAi construct with ethanol. You will analyse the consequences of ADF inhibition on actin organisation and dynamics. In addition you will analyse the consequences of ADF inhibition on plant development. These experiments will give insight in the role of ADF on actin organisation and dynamics in live cells. Moreover, this study will reveal the consequences of altered actin configuration and dynamics on cell growth and plant development.
Used skills Ethanol inducible RNAi, Confocal microscopy, Computer assisted data analysis
Requirements PCB 30306; PPH 30306 (advised, not obliged)