Paraquat induces actin assembly in depolymerizing conditions.
The molecular mechanism (or mechanisms) at the basis of paraquat (PQ) (a widely used herbicide) toxicity is far from being fully understood. Until now, two main points of view have emerged: 1) PQ-related cell injuries could be mediated by toxic oxygen free radicals coming from the metabolism of the herbicide by the microsomal enzyme system, and/or 2) PQ, by inducing mitochondrial swelling and breakage, could cause troubles in cell energy charge, then driving the cell to death. Recently, some of cytoskeletal structures (microtubules and microfilaments) have been proposed as further PQ cell targets. The microfilament system in particular seems to be markedly affected by the herbicide, but so far no direct evidence associates PQ to actin damage. In this study, experimental data are presented concerning the direct effect of PQ on actin dynamics in solution. We demonstrate that actin selectively binds PQ; moreover, PQ induces the formation of actin sopramolecular structures in depolymerizing medium (G-buffer). Furthermore, by the interactions with F-actin cross-linking proteins (alpha-actinin and filamin), FITC-phalloidin, and myosin subfragment 1 (S1), it is demonstrated that PQ-induced actin aggregates are undoubtedly built up by F-actin. Electron micrographs showed that PQ-induced actin polymers are very short and tend to aggregate one to another. This mutual cohesion leads to the steric blockage of polymer growing ends as suggested by nucleated actin polymerization assays. Sonication, by releasing F-actin fragments from short polymer aggregates, allows actin polymer ends to regain their growing ability.
Milzani
A
A
Department of Biology, University of Milan, Italy.
Dalledonne
I
I
Vailati
G
G
Colombo
R
R
D000185:Actinin / Q000378:metabolism; D000199:Actins / Q000187:drug effects* / Q000378:metabolism; D003432:Cross-Linking Reagents; D006540:Herbicides / Q000378:metabolism / Q000494:pharmacology*; D008840:Microfilament Proteins / Q000378:metabolism*; D009218:Myosins / Q000378:metabolism; D010269:Paraquat / Q000378:metabolism / Q000494:pharmacology*; D010449:Peptide Mapping; D010590:Phalloidine / Q000378:metabolism; D011108:Polymers; D012542:Scattering, Radiation; D012996:Solutions; D013050:Spectrometry, Fluorescence; D014783:Viscosity
D000199:Actins; D003432:Cross-Linking Reagents; D006540:Herbicides; D008840:Microfilament Proteins; D011108:Polymers; D012996:Solutions; D000185:Actinin; D010590:Phalloidine; D009218:Myosins; D010269:Paraquat