Brown, LA, O'Leary-Steele, C, Brookes, P et al. (4 more authors) (2011) A small molecule with differential effects on the PTS1 and PTS2 peroxisome matrix import pathways. The Plant Journal, 65 (6). 980 - 990. ISSN 0960-7412
Abstract
The use of small molecules has great power to dissect biological processes. This study presents the identification and characterisation of an inhibitor of peroxisome matrix protein import. A mini-screen was carried out to identify molecules that cause alteration in peroxisome morphology, or mislocalization of a peroxisome targeted fluorescent reporter protein. A benzimidazole lead compound (LDS-003655) was identified that resulted in reduced GFP fluorescence in peroxisomes and cytosolic GFP accumulation. The effect of the compound was specific to peroxisomes as Golgi bodies, endoplasmic reticulum and the actin cytoskeleton were unaffected even at 25 μm, whereas peroxisome import via the PTS1 pathway was compromised at 100 nm. When seedlings were grown on 25 μm LDS-003655 they displayed morphology typical of seedlings grown in the presence of auxin, and expression of the auxin reporter DR5::GFP was induced. Analysis of a focussed library of LDS-003655 derivatives in comparison with known auxins led to the conclusion that the auxin-like activity of LDS-003655 is attributable to its in situ hydrolysis giving rise to 2,5-dichlorobenzoic acid, whereas the import inhibiting activity of LDS-003655 requires the whole molecule. None of the auxins tested had any effect on peroxisome protein import. Matrix import by the PTS2 import pathway was relatively insensitive to LDS-003655 and its active analogues, with effects only seen after prolonged incubation on high concentrations. Steady-state protein levels of PEX5, the PTS1 import pathway receptor, were reduced in the presence of 100 nm LDS-003655, suggesting a possible mechanism for the import inhibition.
Metadata
Item Type: | Article |
---|---|
Authors/Creators: |
|
Copyright, Publisher and Additional Information: | (c) 2011 The Authors The Plant Journal (c) 2011 Blackwell Publishing Ltd. This is the pre-peer reviewed version of the following article: Brown, LA, O'Leary-Steele, C, Brookes, P, Armitage, L, Kepinski, S, Warriner, SL and Baker, A (2011) A small molecule with differential effects on the PTS1 and PTS2 peroxisome matrix import pathways. The Plant Journal, 65 (6). 980 - 990. ISSN 0960-7412, which has been published in final form at http://dx.doi.org/10.1111/j.1365-313X.2010.04473.x Uploaded in accordance with the publisher's self-archiving policy |
Keywords: | peroxisome; chemical genetics; receptor; protein import; auxin; endoplasmic-reticulum; arabidopsis-thaliana; chemical genetics; golgi-apparatus; receptor; proteins; plants; PEX5P; transport; germination |
Dates: |
|
Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Chemistry (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 29 Jan 2014 16:30 |
Last Modified: | 16 Nov 2016 09:49 |
Published Version: | http://dx.doi.org/10.1111/j.1365-313X.2010.04473.x |
Status: | Published |
Publisher: | Wiley-Blackwell |
Identification Number: | 10.1111/j.1365-313X.2010.04473.x |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:77544 |