Journal Article

Time-course of changes in amounts of specific proteins upon exposure to hyper-<i>g,</i> 2-D clinorotation, and 3-D random positioning of <i>Arabidopsis</i> cell cultures

Žarko Barjaktarović, Alfred Nordheim, Tobias Lamkemeyer, Claudia Fladerer, Johannes Madlung and Rüdiger Hampp

in Journal of Experimental Botany

Published on behalf of Society for Experimental Biology

Volume 58, issue 15-16, pages 4357-4363
Published in print December 2007 | ISSN: 0022-0957
Published online December 2007 | e-ISSN: 1460-2431 | DOI:

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In previous studies it has been shown that callus cell cultures of Arabidopsis thaliana respond to changes in gravitational field strengths by altered gene expression. In this study an investigation was carried out into how different g conditions affect the proteome of such cells. For this purpose, callus cells were exposed to 8 g (centrifugation) and simulated microgravity (2-D clinorotation: fast rotating clinostat, yielding 0.0016 g at maximum; and 3-D random positioning) for up to 16 h. Extracts containing total soluble protein were subjected to 2-D SDS–PAGE. Image analysis of Sypro Ruby®-stained gels showed that ∼28 spots reproducibly and significantly (P <0.05) changed in amount after 2 h of hypergravity (18 up- and 10 down-regulated). These spots were analysed by electrospray ionization tandem mass spectrometry (ESI-MS/MS). In the case of 2-D clinorotation, 19 proteins changed in a manner similar to hypergravity, while random positioning affected only eight spots. Identified proteins were mainly stress related, and are involved in detoxification of reactive oxygen species, signalling, and calcium binding. Surprisingly, centrifugation and clinorotation showed homologies which were not detected for random positioning. The data indicate that simulation of weightlessness is different between clinorotation and random positioning.

Keywords: Arabidopsis thaliana; cell cultures; hypergravity; proteomics; simulated microgravity

Journal Article.  4014 words.  Illustrated.

Subjects: Plant Sciences and Forestry

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