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Nuclear bodies and compartmentalization of pre-mRNA splicing factors in higher plants

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Abstract

We studied the fine structural organization of nuclear bodies in the root meristem during germination of maize and Arabidopsis thaliana using electron microscopy (EM). Cajal bodies (CBs) were observed in quiescent embryos and germinating cells in both species. The number and distribution of CBs were investigated. To characterize the nuclear splicing domains, immunofluorescence labelling with antibodies against splicing factors (U2B″ and m3G-snRNAs) and in situ hybridisation (with U1/U6 antisense probes) were performed combined with confocal microscopy. Antibodies specific to the Arabidopsis SR splicing factor atRSp31 were produced. AtRSp31 was detected in quiescent nuclei and in germinating cells. This study revealed an unexpected speckled nuclear organization of atRSp31 in root epidermal cells where micro-clusters of interchromatin granules were also observed by EM. Therefore, we examined the distribution of green fluorescent protein (GFP)-tagged atRSp31 in living cells after Agrobacterium -mediated transient expression. When expressed transiently, atRSp31-GFP exhibited a speckled distribution in leaf cells. Treatments with α-amanitin, okadaic acid, staurosporine or heat shock induced the speckles to reorganize. Furthermore, we generated stable Arabidopsis transgenics expressing atRSp31-GFP. The distribution of the fusion protein was identical to that of endogenous atRSp31. Three-dimensional time-lapse confocal microscopy showed that speckles were highly dynamic domains over time.

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Acknowledgements

We are grateful to Dr P. Gérard (Université de Liège; Unité de Recherche de Statistique — Aspects Expérimentaux) for assistance with statistical analysis, Prof. Marc Boutry (Université Catholique de Louvain, Belgium) for valuable advice with transient transformation and to Prof. Peter Shaw (John Innes Centre, Norwich, UK) for helpful comments. This research was supported by grants from “National Fund for Scientific Research” (grants no. 2.4547.99, 2.4520.02 and 2.4542.00) and from “Fonds Spéciaux du Conseil de la Recherche” of the University of Liège. S.D. was supported by an FRIA grant (Fonds de la Recherche pour l’Industrie et l’Agriculture) and V.T. is supported by FRIA.

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  1. Laboratory of Plant Cell Biology, Department of Life Sciences, University of Liège, Bld du Rectorat, 27 Sart Tilman, 4000 Liège, Belgium

    Sarah Docquier, Vinciane Tillemans, Roger Deltour & Patrick Motte

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  1. Sarah Docquier
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  2. Vinciane Tillemans
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  3. Roger Deltour
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  4. Patrick Motte
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Correspondence to Patrick Motte.

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Communicated by P. Shaw

S. Docquier and P. Motte contributed equally to this work

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Docquier, S., Tillemans, V., Deltour, R. et al. Nuclear bodies and compartmentalization of pre-mRNA splicing factors in higher plants. Chromosoma 112, 255–266 (2004). https://doi.org/10.1007/s00412-003-0271-3

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  • DOI: https://doi.org/10.1007/s00412-003-0271-3

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