Functional distribution and dynamics of Arabidopsis SR splicing factors in living plant cells

[en] Serine/arginine-rich (SR) proteins constitute an important class of splicing regulators in higher eukaryotes that share a modular structure consisting of one or two N-terminal RNA recognition motif (RRM) domains and a C-terminal RS-rich domain. Herein, we have investigated the in vivo functional distribution of Arabidopsis SR factors. Agrobacterium-mediated transient transformation revealed nuclear speckled distribution and the overall colocalization of fluorescent protein (FP)-tagged SR factors in both tobacco and Arabidopsis cells. Their overall colocalization in larger nucleoplasmic domains was further observed after transcriptional and phosphorylation/dephosphorylation inhibition, indicating a close functional association between SR factors, independent of their phosphorylation state. Furthermore, we demonstrated in vivo the conserved role of the RS and RRM domains in the efficient targeting of Arabidopsis SR proteins to nuclear speckles by using a series of structural domain-deleted mutants of atRSp31 and atRSZp22. We suggest additional roles of RS domain such as the shuttling of atRSZp22 between nucleoplasm and nucleolus through its phosphorylation level. The coexpression of deletion mutants with wild-type SR proteins revealed potential complex associations between them. Fluorescence recovery after photobleaching demonstrated similar dynamic properties of SR factors in both tobacco transiently expressing cells and Arabidopsis transgenics. Cell cycle phase-dependent organization of FP-tagged SR proteins was observed in living tobacco BY-2 cells. We showed that atRSp31 is degraded at metaphase by fluorescence quantification. SR proteins also localized within small foci at anaphase. These results demonstrate interesting related features as well as potentially important differences between plant and animal SR proteins.

Disciplines :

Phytobiology (plant sciences, forestry, mycology...)

Author, co-author :

Tillemans, Vinciane ; Université de Liège - ULiège > Département des sciences de la vie > Génomique fonctionnelle et imagerie moléculaire végétale

Dispa, Laurence ; Université de Liège - ULiège > Biologie cellulaire végétale

Remacle, Claire ; Université de Liège - ULiège > Département des sciences de la vie > Génétique

Collinge, M.

Motte, Patrick ; Université de Liège - ULiège > Département des sciences de la vie > Génomique fonctionnelle et imagerie moléculaire végétale

Language :

English

Title :

Functional distribution and dynamics of Arabidopsis SR splicing factors in living plant cells

Publication date :

February 2005

Journal title :

Plant Journal

ISSN :

0960-7412

eISSN :

1365-313X

Publisher :

Blackwell, Oxford, United Kingdom

Volume :

Issue :

Pages :

567-582

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 10 March 2009

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99 (15 by ULiège)

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156 (12 by ULiège)

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