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JXB Advance Access published online on October 27, 2009
Journal of Experimental Botany, doi:10.1093/jxb/erp315
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© The Author [2009]. 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

RESEARCH PAPER

Golgi membrane dynamics after induction of a dominant-negative mutant Sar1 GTPase in tobacco

Anne Osterrieder, Eric Hummel *, Claudine M. Carvalho {dagger} and Chris Hawes{ddagger}

School of Life Sciences, Oxford Brookes University, Headington, Oxford OX3 0BP, UK

{ddagger} To whom correspondence should be addressed. E-mail: chawes{at}brookes.ac.uk

An inducible system has been established in Nicotiana tabacum plants allowing controlled expression of Sar1-GTP and thus the investigation of protein dynamics after inhibition of endoplasmic reticulum (ER) to Golgi transport. Complete Golgi disassembly and redistribution of Golgi markers into the ER was observed within 18–24h after induction. At the ultrastructural level Sar1-GTP expression led to a decrease in Golgi stack size followed by Golgi fragmentation and accumulation of vesicle remnants. Induction of Sar1-GTP resulted in redistribution of the green fluorescent protein (GFP)-tagged Arabidopsis golgins AtCASP and GC1 (golgin candidate 1, an Arabidopsis golgin 84 isoform) into the ER or cytoplasm, respectively. Additionally, both fusion proteins were observed in punctate structures, which co-located with a yellow fluorescent protein (YFP)-tagged version of Sar1-GTP. The Sar1-GTP-inducible system is compared with constitutive Sar1-GTP expression and brefeldin A treatment, and its potential for the study of the composition of ER exit sites and early cis-Golgi structures is discussed.

Key words: Brefeldin A, ER exit sites, Golgi apparatus, golgins, inducible system, matrix, Sar1, Sar1-GTP

* Present address: Universität Bayreuth, Elektronenmikroskopie NW I/B1, 95440 Bayreuth, Germany.

{dagger} Present address: Department of Plant Pathology, Federal University of Viçosa, 36570000 Viçosa-MG, Brazil.

Received 13 August 2009; Revised 25 September 2009 Accepted 5 October 2009


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