Skip Navigation


Plant and Cell Physiology Advance Access originally published online on July 30, 2008
Plant and Cell Physiology 2008 49(9):1331-1341; doi:10.1093/pcp/pcn108
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
49/9/1331    most recent
pcn108v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by Eudes, A.
Right arrow Articles by Jouanin, L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Eudes, A.
Right arrow Articles by Jouanin, L.
Agricola
Right arrow Articles by Eudes, A.
Right arrow Articles by Jouanin, L.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Purification, Cloning and Functional Characterization of an Endogenous beta-Glucuronidase in Arabidopsis thaliana

Aymerick Eudes1,3, Grégory Mouille1, Johanne Thévenin1, Arnaud Goyallon2, Zoran Minic1 and Lise Jouanin1,*

1 INRA, Centre de Versailles, Institut Jean-Pierre Bourgin, Laboratoire de Biologie Cellulaire, 78026 Versailles cedex, France
2 INRA, Centre de Versailles, Institut Jean-Pierre Bourgin, Plateforme de Chimie du Végétal, 78026 Versailles cedex, France

*Corresponding author: E-mail, jouanin{at}versailles.inra.fr; Fax, +33-1-30-83-30-99.


  Abstract

Beta-glucuronidase (GUS) activities have been extensively characterized in bacteria, fungi, and animals, and the bacterial enzyme GUSA from Escherichia coli is commonly used as a reporter for gene expression studies in plants. Although endogenous GUS activity has been observed in plants, the nature and function of the enzymes involved remain elusive. Here we report on tissue-specific localization, partial purification and identification of AtGUS2, a GUS active under acidic conditions from Arabidopsis thaliana. This enzyme belongs to the GH79 family in the Carbohydrate-Active Enzymes database, which also includes mammalian heparanases that degrade the carbohydrate moieties of cell surface proteoglycans, and fungal enzymes active on arabinogalactan proteins (AGPs). We characterized a knockout insertion line (atgus2-1) and transgenic lines overexpressing AtGUS2 (Pro35S:AtGUS2). Endogenous GUS activity assayed histochemically and biochemically was absent in atgus2-1 tissues and four times higher in Pro35S:AtGUS2 lines. AGPs purified from atgus2-1 and Pro35S:AtGUS2 seedlings showed higher and markedly lower glucuronic acid content, respectively. Our results suggest that endogenous GUS activity influences the sugar composition of the complex polysaccharide chains of AGPs. We also show that transgenics display hypocotyl and root growth defects compared to wild-type plants. Hypocotyl and root lengths are increased in Pro35S:AtGUS2 seedlings, whereas hypocoyl length is reduced in atgus2-1 seedlings. These data are consistent with a role for the carbohydrate moieties of AGPs in cell growth.

Keywords: Arabidopsis - Arabinogalactan protein - Beta-glucuronidase - Family 79 glycoside hydrolase - Hypocotyl and root growth

Abbreviations: AGPs, arabinogalactan proteins; ConA, concanavalin A; GH, glycoside hydrolase; GlcA, glucuronic acid; GUS, beta-glucuronidase; MALDI-TOF MS, matrix assisted laser-desorption ionization time-of-flight mass spectrometry; pNPGlcA, p-nitrophenyl-β-D-glucuronide; X-GlcA, 5-bromo-4-chloro-3-β-indolyl-glucuronide.

3Present address: University of Florida, Horticultural Sciences Department, PO Box 110690, Gainesville, FL 32611, USA.

(Received July 8, 2008; Accepted July 27, 2008)
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.