title: Hexavalent chromium-induced differential disruption of cortical microtubules in some Fabaceae species is correlated with acetylation of α-tubulin.
abstract: The effects of hexavalent chromium [Cr(VI)] on the cortical microtubules (MTs) of five species of the Fabaceae family (Vicia faba, Pisum sativum, Vigna sinensis, Vigna angularis, and Medicago sativa) were investigated by confocal laser scanning microscopy after immunolocalization of total tubulin with conventional immunofluorescence techniques and of acetylated α-tubulin with the specific 6-11B-1 monoclonal antibody. Moreover, total α-tubulin and acetylated α-tubulin were quantified by Western immunoblotting and scanning densitometry. Results showed the universality of Cr(VI) detrimental effects to cortical MTs, which proved to be a sensitive and reliable subcellular marker for monitoring Cr(VI) toxicity in plant cells. However, a species-specific response was recorded, and a correlation of MT disturbance with the acetylation status of α-tubulin was demonstrated. In V. faba, MTs were depolymerized at the gain of cytoplasmic tubulin background and displayed low α-tubulin acetylation, while in P. sativum, V. sinensis, V. angularis, and M. sativa, MTs became bundled and changed orientation from perpendicular to oblique or longitudinal. Bundled MTs were highly acetylated as determined by both immunofluorescence and Western immunoblotting. Tubulin acetylation in P. sativum and M. sativa preceded MT bundling; in V. sinensis it followed MT derangement, while in V. angularis the two phenomena coincided. Total α-tubulin remained constant in all treatments. Should acetylation be an indicator of MT stabilization, it is deduced that bundled MTs became stabilized, lost their dynamic properties, and were rendered inactive. Results of this report allow the conclusion that Cr(VI) toxicity disrupts MTs and deranges the MT-mediated functions either by depolymerizing or stabilizing them.
authors.affiliation: Department of Botany, School of Biology, Aristotle University of Thessaloniki, 541 24, Thessaloniki, Greece. eelefth@bio.auth.gr.
authors.forename: Eleftherios P
authors.initials: EP
authors.lastname: Eleftheriou
authors.affiliation: Department of Botany, School of Biology, Aristotle University of Thessaloniki, 541 24, Thessaloniki, Greece.
authors.forename: Ioannis-Dimosthenis S
authors.initials: ID
authors.lastname: Adamakis
authors.affiliation: Department of Botany, School of Biology, Aristotle University of Thessaloniki, 541 24, Thessaloniki, Greece.
authors.forename: Vasiliki A
authors.initials: VA
authors.lastname: Michalopoulou
mesh_terms: D000107:Acetylation; D002857:Chromium / Q000494:pharmacology*; D004785:Environmental Pollutants / Q000494:pharmacology*; D007887:Fabaceae / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008870:Microtubules; D010940:Plant Proteins / Q000378:metabolism*; D018517:Plant Roots / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D014404:Tubulin / Q000378:metabolism*; D050257:Tubulin Modulators / Q000494:pharmacology
chemical_list: D004785:Environmental Pollutants; D010940:Plant Proteins; D014404:Tubulin; D050257:Tubulin Modulators; D002857:Chromium; C074702:chromium hexavalent ion
keywords: Cortical microtubules; Depolymerization; Fabaceae; Hexavalent chromium; Stabilization; Tubulin acetylation