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α-amylase is not required for breakdown of transitory starch in Arabidopsis leaves

Citation

Yu, T-S and Zeeman, SC and Thorneycroft, D and Fulton, DC and Dunstan, H and Lue, W-L and Hegemann, B and Tung, S-Y and Umemoto, T and Chapple, A and Tsai, D-L and Wang, S-M and Smith, AM and Chen, J and Smith, SM, α-amylase is not required for breakdown of transitory starch in Arabidopsis leaves, Journal of Biological Chemistry, 280, (11) pp. 9773-9779. ISSN 0021-9258 (2005) [Refereed Article]

Copyright Statement

2005 by The American Society for Biochemistry and Molecular Biology

DOI: doi:10.1074/jbc.M413638200

Abstract

The Arabidopsis thaliana genome encodes three α-amylase-like proteins (AtAMY1, AtAMY2, and AtAMY3). Only AtAMY3 has a predicted N-terminal transit peptide for plastidial localization. AtAMY3 is an unusually large α-amylase (93.5 kDa) with the C-terminal half showing similarity to other known α-amylases. When expressed in Escherichia coli, both the whole AtAMY3 protein and the C-terminal half alone show α-amylase activity. We show that AtAMY3 is localized in chloroplasts. The starch-excess mutant of Arabidopsis sex4, previously shown to have reduced plastidial α-amylase activity, is deficient in AtAMY3 protein. Unexpectedly, T-DNA knock-out mutants of AtAMY3 have the same diurnal pattern of transitory starch metabolism as the wild type. These results show that AtAMY3 is not required for transitory starch breakdown and that the starch-excess phenotype of the sex4 mutant is not caused simply by deficiency of AtAMY3 protein. Knock-out mutants in the predicted non-plastidial α-amylases AtAMY1 and AtAMY2 were also isolated, and these displayed normal starch breakdown in the dark as expected for extraplastidial amylases. Furthermore, all three AtAMY double knock-out mutant combinations and the triple knock-out degraded their leaf starch normally. We conclude that α-amylase is not necessary for transitory starch breakdown in Arabidopsis leaves.

Item Details

Item Type:Refereed Article
Keywords:arabidopsis thaliana, alpha-amylase proteins, starch breakdown
Research Division:Biological Sciences
Research Group:Plant Biology
Research Field:Plant Physiology
Objective Division:Expanding Knowledge
Objective Group:Expanding Knowledge
Objective Field:Expanding Knowledge in the Biological Sciences
Author:Smith, SM (Professor Steven Smith)
ID Code:101519
Year Published:2005
Web of Science® Times Cited:100
Deposited By:Plant Science
Deposited On:2015-06-25
Last Modified:2015-09-25
Downloads:0

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