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The cell wall of Arabidopsis thaliana influences actin network dynamics

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Tolmie, Frances, Poulet, Axel, McKenna, Joseph F., Sassmann, Stefan, Graumann, Katja, Deeks, Michael, Runions, John and Bozhkov, Peter (2017) The cell wall of Arabidopsis thaliana influences actin network dynamics. Journal of Experimental Botany, 68 (16). pp. 4517-4527. doi:10.1093/jxb/erx269

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Official URL: http://dx.doi.org/10.1093/jxb/erx269

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Abstract

In plant cells, molecular connections link the cell wall–plasma membrane–actin cytoskeleton to form a continuum. It is hypothesized that the cell wall provides stable anchor points around which the actin cytoskeleton remodels. Here we use live cell imaging of fluorescently labelled marker proteins to quantify the organization and dynamics of the actin cytoskeleton and to determine the impact of disrupting connections within the continuum. Labelling of the actin cytoskeleton with green fluorescent protein (GFP)–fimbrin actin-binding domain 2 (FABD2) resulted in a network composed of fine filaments and thicker bundles that appeared as a highly dynamic remodelling meshwork. This differed substantially from the GFP–Lifeact-labelled network that appeared much more sparse with thick bundles that underwent ‘simple movement’, in which the bundles slightly change position, but in such a manner that the structure of the network was not substantially altered during the time of observation. Label-dependent differences in actin network morphology and remodelling necessitated development of two new image analysis techniques. The first of these, ‘pairwise image subtraction’, was applied to measurement of the more rapidly remodelling actin network labelled with GFP–FABD2, while the second, ‘cumulative fluorescence intensity’, was used to measure bulk remodelling of the actin cytoskeleton when labelled with GFP–Lifeact. In each case, these analysis techniques show that the actin cytoskeleton has a decreased rate of bulk remodelling when the cell wall–plasma membrane–actin continuum is disrupted either by plasmolysis or with isoxaben, a drug that specifically inhibits cellulose deposition. Changes in the rate of actin remodelling also affect its functionality, as observed by alteration in Golgi body motility.

Item Type: Journal Article
Divisions: Faculty of Science > Life Sciences (2010- )
Journal or Publication Title: Journal of Experimental Botany
Publisher: Oxford University Press
ISSN: 0022-0957
Official Date: 3 October 2017
Dates:
DateEvent
3 October 2017Published
Volume: 68
Number: 16
Page Range: pp. 4517-4527
DOI: 10.1093/jxb/erx269
Status: Peer Reviewed
Publication Status: Published
Access rights to Published version: Restricted or Subscription Access

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