Journal Article

Localization and Topogenesis Studies of Cytoplasmic and Vacuolar Homologs of the <i>Galanthus nivalis</i> Agglutinin

Elke Fouquaert, Sally L. Hanton, Federica Brandizzi, Willy J. Peumans and Els J. M. Van Damme

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 48, issue 7, pages 1010-1021
Published in print July 2007 | ISSN: 0032-0781
Published online July 2007 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pcm071
Localization and Topogenesis Studies of Cytoplasmic and Vacuolar Homologs of the Galanthus nivalis Agglutinin

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  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

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The Galanthus nivalis agglutinin (GNA) is synthesized as a preproprotein. To corroborate the role of the different targeting peptides in the topogenesis of GNA and related proteins, different constructs were made whereby both the complete original GNA gene and different truncated sequences were coupled to the enhanced green fluorescent protein (EGFP). In addition, a GNA ortholog from rice that lacks the signal peptide and C-terminal propeptide sequence was fused to EGFP. These fusion constructs were expressed in tobacco BY-2 cells and their localization analyzed by confocal fluorescence microscopy. We observed that the processed preproprotein of GNA was directed towards the vacuolar compartment, whereas both the truncated forms of GNA corresponding to the mature lectin polypeptide and the rice ortholog of GNA were located in the nucleus and the cytoplasm. It can be concluded, therefore, that removal of the C-terminal propeptide and the signal peptide is sufficient to change the subcellular targeting of a normally vacuolar protein to the nuclear/cytoplasmic compartment of the BY-2 cells. These findings support the proposed hypothesis that cytoplasmic/nuclear GNA-like proteins and their vacuolar homologs are evolutionarily related and that the classical GNA-related lectins might have evolved from cytoplasmic orthologs through an evolutionary event involving the insertion of a signal peptide and a C-terminal propeptide.

Keywords: Cytoplasmic ortholog; Galanthus nivalis agglutinin; Lectin; Subcellular location

Journal Article.  6639 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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