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Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
RESEARCH ARTICLE

Arabidopsis phospholipase Dδ as an initiator of cytoskeleton-mediated signalling to fundamental cellular processes

Angela Y. Y. Ho A , David A. Day A , Melissa H. Brown A B and Jan Marc A C
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A School of Biological Sciences, Macleay Building A12, University of Sydney, Sydney, NSW 2006, Australia.

B Present address: School of Biological Sciences, Flinders University, Adelaide, SA 5042, Australia.

C Corresponding author. Email: jmarc@bio.usyd.edu.au

Functional Plant Biology 36(2) 190-198 https://doi.org/10.1071/FP08222
Submitted: 15 August 2008  Accepted: 10 December 2008   Published: 5 February 2009

Abstract

Phospholipase D (PLD), in combination with the cytoskeleton, plays a key role in plant signal transduction. One isotype of the multigene Arabidopsis PLD family, AtPLDδ, has been implicated in binding microtubules, although the molecular details of the mechanism and identities of potential interaction partners are unclear. We constructed a GFP-AtPLDδ reporter gene, stably transformed it into an Arabidopsis suspension cell line, and used epitope-tagged affinity pull-down assays to isolate a complex of co-purifying proteins. Mass spectrometry analysis of the complex revealed a set of proteins including β-tubulin, actin 7, HSP70, clathrin heavy chain, ATP synthase subunits, and a band 7–4/flotillin homologue. Sequence alignments with defined tubulin- and actin-binding regions from human HsPLD2 revealed highly homologous regions in all 12 AtPLD isotypes, suggesting direct interactions of AtPLDδ with tubulin and actin, while interactions with the remaining partners are likely to be mediated by the cytoskeleton. We propose that AtPLDδ acts through a complex of cytoskeletal and partner proteins to modulate fundamental cellular processes such as cytoskeletal rearrangements, vesicular trafficking, assembly of Golgi apparatus, mitosis and cytokinesis.

Additional keywords: actin, clathrin, environmental stress, heat shock protein, signal transduction, tubulin.


Acknowledgements

We thank Dr Ben Crossett (University of Sydney) for help with mass spectrometry, and Professor Harvey Millar (University of Western Australia) for providing the Arabidopsis cell culture. This work was supported by a Grains Research and Development Corporation scholarship to A.H. and Australian Research Council grant DP0453114 to J.M.


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