Tobacco Arp3 is localized to actin-nucleating sites in vivo

The polarity of actin is a central determinant of intracellular transport in plant cells. To visualize actin polarity in living plant cells, the tobacco homologue of the actin-related protein 3 (ARP3) was cloned and a fusion with the red fluorescent protein (RFP) was generated. Upon transient expres...

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Bibliographic Details
Published in:Journal of experimental botany 2009-02, Vol.60 (2), p.603-614
Main Authors: Maisch, Jan, Fišerová, Jindřiška, Fischer, Lukáš, Nick, Peter
Format: Article
Language:English
Subjects:
Actin
Actin Cytoskeleton - metabolism
actin-related protein 3 (ARP3)
Actin-Related Protein 3 - chemistry
Actin-Related Protein 3 - isolation & purification
Actin-Related Protein 3 - metabolism
Actins
Actins - metabolism
Amino Acid Sequence
Animal cells
Auxins
Biological and medical sciences
Cell Division
Cell lines
Cell Nucleus - metabolism
Cells
Fluorescence
Fundamental and applied biological sciences. Psychology
Green Fluorescent Proteins - metabolism
Luminescent Proteins - metabolism
Membrane Glycoproteins - metabolism
Microfilament Proteins - metabolism
Microfilaments
Molecular Sequence Data
Nicotiana - cytology
Nicotiana - metabolism
Nucleation
Phylogeny
Plant cells
Plant Proteins - metabolism
Plants
Protein Transport
Recombinant Fusion Proteins - metabolism
Red Fluorescent Protein
RESEARCH PAPER
Research Papers
tobacco BY-2
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Summary:The polarity of actin is a central determinant of intracellular transport in plant cells. To visualize actin polarity in living plant cells, the tobacco homologue of the actin-related protein 3 (ARP3) was cloned and a fusion with the red fluorescent protein (RFP) was generated. Upon transient expression of these fusions in the tobacco cell line BY-2 (Nicotiana tabacum L. cv. Bright Yellow 2), punctate structures were observed near the nuclear envelope and in the cortical plasma. These dots could be shown to decorate actin filaments by expressing RFP-ARP3 in a marker line, where actin was tagged by GFP (green fluorescent protein)-FABD (fimbrin actin-binding domain 2). When actin filaments were disrupted by latrunculin B or by prolonged cold treatment, and subsequently allowed to recover, the actin filaments reformed from the RFP-ARP3 structures, that therefore represented actin nucleation sites. The intracellular distribution of these sites was followed during the formation of pluricellular files, and it was observed that the density of RFP-ARP3 increased in the apex of the polarized, terminal cells of a file, whereas it was equally distributed in the central cells of a file. These findings are interpreted in terms of position-dependent differences of actin organization.
ISSN:0022-0957
1460-2431
DOI:10.1093/jxb/ern307