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The Arabidopsis thaliana GSK3/Shaggy like kinase AtSK3-2 modulates floral cell expansion

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Abstract

The GSK3/Shaggy family of serine/threonine protein kinases is involved in a series of biological processes in animals, plants and yeast [Charrier et al. (2002) Plant Physiol 130:577–590; Jope and Johnson (2004) Trends Biochem Sci 29:95–102; Li and Nam (2002) Science 295:1299–1301; Piao et al. (2001) Plant J 27:305–314]. In Arabidopsis thaliana, out of the 10 members of the GSK3/Shaggy-like gene family (AtSKs), a biological function has been assigned to only 1 member (AtSK2-1) by mutation. In the present work, a study was undertaken to elucidate the function of AtSK3-2. We have generated mutated versions of the A. thaliana Shaggy-like kinase 3-2 (AtSK3-2), in which Lys167 and Arg178, respectively homologus to Lys85 and Arg96 of the mammal GSK3β, were modified into Ala by site-directed mutagenesis. In vitro kinase activity assays of the mutated recombinant protein AtSK3-2-R178A showed that the “primed activity” of the mutated kinase was reduced by 90% while the “non-primed” activity was only 20% reduced compared to the wild-type protein kinase. However, the mutant protein AtSK3-2-K167A showed no activity. Arabidopsis transgenic lines over-expressing AtSK3-2-R178A displayed smaller floral organs, namely pedicels, sepals and petals. Conversely, over-expression of both the wild-type AtSK3-2 protein and the AtSK3-2-K167A mutated version, displayed no altered morphogenesis. Scanning electron microscopic analyses of the AtSK3-2-R178A transgenic plants clearly showed a reduced cell size in flower organs, in which quantitative RT-PCR expression analyses of cell wall expansion enzymes showed reduced transcript levels of three xyloglucan endotransglycosylases (XET), namely XTH22 (TCH4), XTH23 (XTR6) and XTH30 (XTR4). Our data show that AtSK3-2 plays an important role in the control of cell elongation in flower development.

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Acknowledgements

We thank A. Picaud for his assistance in the biochemistry experiments and P. Ratet (ISV, Gif/Yvette) for providing the plasmid pCP60. We are grateful to S. Domenichini (IBP, Université Paris-Sud) for SEM expertise, to R. Boyer (IBP, Université Paris-Sud) for photographic work and to Y. Deveaux for advice and fruitful suggestions.

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  1. Bénédicte Charrier

    Present address: Station Biologique, UMR 7139, BP-74, Roscoff, 29682, France

Authors and Affiliations

  1. Institut de Biotechnologie des Plantes (IBP), UMR CNRS 8618, Laboratoire de Biologie du Développement des Plantes, Université Paris-Sud XI, Bat 630, Orsay Cedex, 91405, France

    Gaëlle Claisse, Bénédicte Charrier & Martin Kreis

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  1. Gaëlle Claisse
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Correspondence to Martin Kreis.

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Claisse, G., Charrier, B. & Kreis, M. The Arabidopsis thaliana GSK3/Shaggy like kinase AtSK3-2 modulates floral cell expansion. Plant Mol Biol 64, 113–124 (2007). https://doi.org/10.1007/s11103-007-9138-y

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