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A nodulin/glutamine synthetase-like fusion protein is implicated in the regulation of root morphogenesis and in signalling triggered by flagellin

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Abstract

The nodulin/glutamine synthetase-like protein (NodGS) that we identified proteomically in Arabidopsis thaliana is a fusion protein composed of an N-terminal amidohydrolase domain that shares homology with nodulins and a C-terminal domain of prokaryotic glutamine synthetase type I. The protein is homologous to the FluG protein, a morphogenetic factor in fungi. Although genes encoding NodGS homologues are present in many plant genomes, their products have not yet been characterized. The Arabidopsis NodGS was present in an oligomeric form of ~700-kDa, mainly in the cytosol, and to a lesser extent in the microsomal membrane fraction. The oligomeric NodGS was incorporated into large heterogeneous protein complexes >700 kDa and partially co-immunoprecipitated with γ-tubulin. In situ and in vivo microscopic analyses revealed a NodGS signal in the cytoplasm, with endomembranes, particularly in the perinuclear area. NodGS had no detectable glutamine synthetase activity. Downregulation of NodGS by RNAi resulted in plants with a short main root, reduced meristematic activity and disrupted development of the root cap. Y2H analysis and publicly available microarray data indicated a role for NodGS in biotic stress signalling. We found that flagellin enhanced the expression of the NodGS protein, which was then preferentially localized in the nuclear periphery. Our results point to a role for NodGS in root morphogenesis and microbial elicitation. These data might help in understanding the family of NodGS/FluG-like fusion genes that are widespread in prokaryotes, fungi and plants.

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Abbreviations

APM:

Amiprophos-methyl

BN-PAGE:

Blue native PAGE

DPX:

Dapoxyl dye

DTT:

Dithiothreitol

ER:

Endoplasmic reticulum

flg22:

Flagellin 22

FRAP:

Fluorescence recovery after photobleaching

GS:

Glutamine synthetase

MS:

Murashige and Skoog

NodGS:

Nodulin/glutamine synthetase-like protein

PMSF:

Phenylmethylsulfonyl fluoride

Y2H:

Yeast two-hybrid analysis

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Acknowledgments

We are thankful to Gabriela Kočárová, Elena Kubátová and Jan Svoboda for technical assistance. We gratefully acknowledge Dr. Csaba Koncz (Max Planck Institute, Cologne) for pAct2 cDNA library, Dr. Roger Tsien (University of California, San Diego, CA, USA) for the kind gift of the binary RFP vector pH7WGR2.0, and Dr. László Bögre (Royal Holloway University of London, UK) for GFP-MAP4 seeds. This work was supported by The Grant Agency of the ASCR grants IAA500200719 and KJB500200705 and, MSMT LC545 project, the Grant Agency of the Czech Republic grant 204/09/H084 and Institutional Research Concept AV0Z50200510.

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  1. Ondřej Plíhal & Beáta Petrovská

    Present address: Centre of the Region Haná for Biotechnological and Agricultural Research, Department of Biochemistry, Faculty of Science, Palacký University in Olomouc, Šlechtitelů 11, 78371, Olomouc, Czech Republic

Authors and Affiliations

  1. Institute of Microbiology, v.v.i., Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220, Prague 4, Czech Republic

    Anna Doskočilová, Ondřej Plíhal, Jindřich Volc, Jana Chumová, Hana Kourová, Petr Halada & Pavla Binarová

  2. Institute of Experimental Botany, v.v.i., Academy of Sciences of the Czech Republic, Sokolovská 6, 77200, Olomouc, Czech Republic

    Beáta Petrovská

  3. Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany AS CR, Sokolovská 6, 77200, Olomouc, Czech Republic

    Beáta Petrovská

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  1. Anna Doskočilová
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Correspondence to Pavla Binarová.

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A. Doskočilová and O. Plíhal contributed equally to this article.

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Doskočilová, A., Plíhal, O., Volc, J. et al. A nodulin/glutamine synthetase-like fusion protein is implicated in the regulation of root morphogenesis and in signalling triggered by flagellin. Planta 234, 459–476 (2011). https://doi.org/10.1007/s00425-011-1419-7

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