Membrane Protein Transport between the Endoplasmic Reticulum and the Golgi in Tobacco Leaves Is Energy Dependent but Cytoskeleton Independent Evidence from Selective Photobleaching

Membrane Protein Transport between the Endoplasmic Reticulum and the Golgi in Tobacco Leaves Is... Federica Brandizzi 1 , a , Erik L. Snapp b , Alison G. Roberts c , Jennifer Lippincott-Schwartz b and Chris Hawes a a Research School of Biological and Molecular Sciences, Oxford Brookes University, Gipsy Lane, Oxford OX3 0BP, United Kingdom b Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892 c Scottish Crop Research Institute, Dundee DD2 5DA, United Kingdom ↵ 1 To whom the correspondence should be addressed. E-mail fbrandizzi@brookes.ac.uk ; fax 44-1865-483955 Abstract The mechanisms that control protein transport between the endoplasmic reticulum (ER) and the Golgi apparatus are poorly characterized in plants. Here, we examine in tobacco leaves the structural relationship between Golgi and ER membranes using electron microscopy and demonstrate that Golgi membranes contain elements that are in close association and/or in direct contact with the ER. We further visualized protein trafficking between the ER and the Golgi using Golgi marker proteins tagged with green fluorescent protein. Using photobleaching techniques, we showed that Golgi membrane markers constitutively cycle to and from the Golgi in an energy-dependent and N -ethylmaleimide–sensitive manner. We found that membrane protein transport toward the Golgi occurs independently of the cytoskeleton and does not require the Golgi to be motile along the surface of the ER. Brefeldin A treatment blocked forward trafficking of Golgi proteins before their redistribution into the ER. Our results indicate that in plant cells, the Golgi apparatus is a dynamic membrane system whose components continuously traffic via membrane trafficking pathways regulated by brefeldin A– and N -ethylmaleimide–sensitive machinery. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png

Membrane Protein Transport between the Endoplasmic Reticulum and the Golgi in Tobacco Leaves Is Energy Dependent but Cytoskeleton Independent Evidence from Selective Photobleaching

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Publisher
American Society of Plant Biologist
Copyright
Copyright © 2015 by the American Society of Plant Biologists
ISSN
1040-4651
eISSN
1532-298X
D.O.I.
10.1105/tpc.001586
Publisher site
See Article on Publisher Site

Abstract

Federica Brandizzi 1 , a , Erik L. Snapp b , Alison G. Roberts c , Jennifer Lippincott-Schwartz b and Chris Hawes a a Research School of Biological and Molecular Sciences, Oxford Brookes University, Gipsy Lane, Oxford OX3 0BP, United Kingdom b Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892 c Scottish Crop Research Institute, Dundee DD2 5DA, United Kingdom ↵ 1 To whom the correspondence should be addressed. E-mail fbrandizzi@brookes.ac.uk ; fax 44-1865-483955 Abstract The mechanisms that control protein transport between the endoplasmic reticulum (ER) and the Golgi apparatus are poorly characterized in plants. Here, we examine in tobacco leaves the structural relationship between Golgi and ER membranes using electron microscopy and demonstrate that Golgi membranes contain elements that are in close association and/or in direct contact with the ER. We further visualized protein trafficking between the ER and the Golgi using Golgi marker proteins tagged with green fluorescent protein. Using photobleaching techniques, we showed that Golgi membrane markers constitutively cycle to and from the Golgi in an energy-dependent and N -ethylmaleimide–sensitive manner. We found that membrane protein transport toward the Golgi occurs independently of the cytoskeleton and does not require the Golgi to be motile along the surface of the ER. Brefeldin A treatment blocked forward trafficking of Golgi proteins before their redistribution into the ER. Our results indicate that in plant cells, the Golgi apparatus is a dynamic membrane system whose components continuously traffic via membrane trafficking pathways regulated by brefeldin A– and N -ethylmaleimide–sensitive machinery.

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