Skip to main content

Research Repository

Advanced Search

Microcompartmentation of cytosolic aldolase by interaction with the actin cytoskeleton in Arabidopsis

Garagounis, Constantine; Kostaki, Kalliopi-Ioanna; Hawkins, Tim J.; Cummins, Ian; Fricker, Mark D.; Hussey, Patrick J.; Hetherington, Alistair M.; Sweetlove, Lee J.

Microcompartmentation of cytosolic aldolase by interaction with the actin cytoskeleton in Arabidopsis Thumbnail


Authors

Constantine Garagounis

Kalliopi-Ioanna Kostaki

Ian Cummins

Mark D. Fricker

Alistair M. Hetherington

Lee J. Sweetlove



Abstract

Evidence is accumulating for molecular microcompartments formed when proteins interact in localized domains with the cytoskeleton, organelle surfaces, and intracellular membranes. To understand the potential functional significance of protein microcompartmentation in plants, we studied the interaction of the glycolytic enzyme fructose bisphosphate aldolase with actin in Arabidopsis thaliana. Homology modelling of a major cytosolic isozyme of aldolase, FBA8, suggested that the tetrameric holoenzyme has two actin binding sites and could therefore act as an actin-bundling protein, as was reported for animal aldolases. This was confirmed by in vitro measurements of an increase in viscosity of F-actin polymerized in the presence of recombinant FBA8. Simultaneously, interaction with F-actin caused non-competitive inhibition of aldolase activity. We did not detect co-localization of an FBA8–RFP fusion protein, expressed in an fba8-knockout background, with the actin cytoskeleton using confocal laser-scanning microscopy. However, we did find evidence for a low level of interaction using FRET-FLIM analysis of FBA8–RFP co-expressed with the actin-binding protein GFP–Lifeact. Furthermore, knockout of FBA8 caused minor alterations of guard cell actin cytoskeleton morphology and resulted in a reduced rate of stomatal closure in response to decreased humidity. We conclude that cytosolic aldolase can be microcompartmented in vivo by interaction with the actin cytoskeleton and may subtly modulate guard cell behaviour as a result.

Journal Article Type Article
Acceptance Date Jan 7, 2017
Online Publication Date Feb 17, 2017
Publication Date Feb 17, 2017
Deposit Date Feb 16, 2017
Publicly Available Date Feb 17, 2018
Journal Journal of Experimental Botany
Print ISSN 0022-0957
Electronic ISSN 1460-2431
Publisher Oxford University Press
Peer Reviewed Peer Reviewed
Volume 68
Issue 5
Pages 885-898
DOI https://doi.org/10.1093/jxb/erx015
Public URL https://durham-repository.worktribe.com/output/1394324

Files

Accepted Journal Article (1.4 Mb)
PDF

Copyright Statement
This is a pre-copyedited, author-produced PDF of an article accepted for publication in Journal of Experimental Botany following peer review. The version of record Garagounis, Constantine, Kostaki, Kalliopi-Ioanna, Hawkins, Tim J., Cummins, Ian, Fricker, Mark D., Hussey, Patrick J., Hetherington, Alistair M. & Sweetlove, Lee J. (2017). Microcompartmentation of cytosolic aldolase by interaction with the actin cytoskeleton in Arabidopsis. Journal Of Experimental Botany 68(5): 885-898 is available online at: https://doi.org/10.1093/jxb/erx015.






You might also like



Downloadable Citations