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JXB Advance Access published online on October 5, 2005
Journal of Experimental Botany, doi:10.1093/jxb/eri289
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© The Author [2005]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org
Received May 31, 2005
Accepted August 15, 2005

The Visible Plant Cell: Biosensors and Bioreceptors Special Issue Article

Photoactivation of GFP reveals protein dynamics within the endoplasmic reticulum membrane

John Runions 1*, Thorsten Brach 2, Sebastian Kühner 2, and Chris Hawes 1

1 Biological and Molecular Sciences, Oxford Brookes University, Oxford OX3 0BP, UK
2 Biological and Molecular Sciences, Oxford Brookes University, Oxford OX3 0BP, UK; Present address: Department of Cell Biology, Heidelberg Institute for Plant Sciences, University of Heidelberg, D-69120 Heidelberg, Germany

* To whom correspondence should be addressed.
John Runions, E-mail: jrunions{at}brookes.ac.uk


  Abstract

Components of the plant cell secretory pathway, including the endoplasmic reticulum and Golgi apparatus, are in constant motion. The photoactivation of GFP has been used to determine that proteins within the membrane of the ER flow as the ER is remodelled. Measurement of the rate at which activated GFP moves away from the activation spot shows that this motion is much faster than would be expected if membrane components moved simply by diffusion. Treatment with latrunculin to depolymerize the actin cytoskeleton stops ER remodelling and reduces the rate of GFP movement to that expected from diffusion alone. This suggests that myosin binds directly or indirectly to ER membrane proteins and actively moves them around over the actin scaffold. Tracking of Golgi body movement was used to demonstrate that they move at the same rate and in the same direction as do photoactivated ER surface proteins. Golgi bodies, therefore, move with, and not over, the surface of the ER. These observations support the current theory of continuity between Golgi bodies and discrete ER exit sites in the ER membrane.

Keywords: Actin cytoskeleton; endoplasmic reticulum; GFP; Golgi; photoactivatable GFP.
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