3-nitrotyrosine and oryzalin

Microtubules (MTs) are essential components of the cytoskeleton and fulfil multiple cellular functions in developmental processes, readily responding to intrinsic and external cues. Nitric oxide signalling is well established in plants, and the MT cytoskeleton is one of its potential targets. To mimic low level nitrosative stress, growth medium was supplemented with 3-nitro-L-tyrosine (NO2-Tyr), a nitrated form of the amino acid tyrosine, and concentration-dependent changes in root growth rate and a reduction in cell division frequencies in Arabidopsis thaliana were observed. In addition, it is reported that exposure to low NO2-Tyr concentrations was not detrimental to plant health and caused subtle and reversible defects. In contrast, growth defects caused by high NO2-Tyr concentrations could not be reversed. Live cell imaging of an MT reporter line revealed that treatment with a low concentration of NO2-Tyr correlated with disorganized cortical MT arrays and associated non-polar cell expansion in the elongation zone. NO2-Tyr treatment antagonized the effects of taxol and oryzalin, further supporting the association of NO2-Tyr with MTs. Furthermore, oblique division plane orientations were observed which were probably induced prior to cytokinesis.

Topics: Arabidopsis; Dinitrobenzenes; Microtubules; Mitosis; Nitric Oxide; Paclitaxel; Plant Roots; Stress, Physiological; Sulfanilamides; Tyrosine

Virtually all eukaryotic alpha-tubulins harbour a C-terminal tyrosine that can be reversibly removed and religated, catalysed by a specific tubulin-tyrosine carboxypeptidase (TTC) and a specific tubulin-tyrosine ligase (TTL), respectively. The biological function of this post-translational modification has remained enigmatic. 3-nitro-L-tyrosine (nitrotyrosine, NO(2)Tyr), can be incorporated into detyrosinated alpha-tubulin instead of tyrosine, producing irreversibly nitrotyrosinated alpha-tubulin. To gain insight into the possible function of detyrosination, the effect of NO(2)Tyr has been assessed in two plant model organisms (rice and tobacco). NO(2)Tyr causes a specific, sensitive, and dose-dependent inhibition of cell division that becomes detectable from 1 h after treatment and which is not observed with non-nitrosylated tyrosine. These effects are most pronounced in cycling tobacco BY-2 cells, where the inhibition of cell division is accompanied by a stimulation of cell length, and a misorientation of cross walls. NO(2)Tyr reduces the abundance of the detyrosinated form of alpha-tubulin whereas the tyrosinated alpha-tubulin is not affected. These findings are discussed with respect to a model where NO(2)Tyr is accepted as substrate by TTL and subsequently blocks TTC activity. The irreversibly tyrosinated alpha-tubulin impairs microtubular functions that are relevant to cell division in general, and cell wall deposition in particular.

Topics: Cell Division; Cell Wall; Dinitrobenzenes; Mitosis; Nicotiana; Oryza; Plant Extracts; Seedlings; Sulfanilamides; Tubulin; Tyrosine