Post-translational modifications of ?-tubulin in Zea mays L. are highly tissue specific

Abstract

To further understand post-translational modifications (PTMs) of plant alpha-tubulin, post-translationally modified alpha-tubulin isoforms from selected tissues of Zea mays L. were examined using two-dimensional electrophoresis and immunoblotting. Except for polyglycylated tubulin, tyrosinated, detyrosinated, acetylated and polyglutamylated alpha-tubulin isoforms were all present in maize tissues. Tyrosinated alpha-tubulin was the predominant variant in all cases, with isoforms alpha1-alpha4 (alpha5) being the most common components. Leaves exhibited a striking difference in PTM patterns of alpha-tubulin isoforms compared to other tissues examined. In leaves, several major specific isoforms were highly modified by detyrosination, acetylation and polyglutamylation. In pollen and anthers, only the most abundant isoform alpha3 was acetylated to an appreciable extent, and no acetylated isoform was found in roots. Similarly, in pollen, anthers and roots, only alpha3 was appreciably polyglutamylated. Additionally, a detyrosinated isoform alpha6 was present in anthers and in leaves, while the tyrosinated isoform alpha6 seemed to be pollen specific. These results indicate that certain types of PTM of plant alpha-tubulin preferentially occur in a tissue-specific way.

Affinity Purification and Characterization of Functional Tubulin from Cell Suspension Cultures of Arabidopsis and Tobacco

“…While polyglutamylationspecific antibodies detected tubulin-like bands by immunoblot and MT-like structures by immunohistochemistry in tobacco and maize (Zea mays) cells (Smertenko et al, 1997;Wang et al, 2004), we obtained no evidence of the polyglutamylation of purified tubulin from Arabidopsis and tobacco cells using the polyglutamylation antibody (Fig. 3) and mass spectrometry (Fig.…”

“…Detyrosination, like acetylation, is generally found on stable and long-lived MTs (Janke, 2014), and kinetochore motors preferentially track detyrosinated MTs during chromosome segregation in animal cell division (Barisic et al, 2015). Previous studies assessed tubulin detyrosination in plant cells using the anti-Glu antibody, which was raised against the animal a-tubulin tail sequence Gly-Glu-Glu-Glu-Gly-Glu-Glu (Smertenko et al, 1997), or by comparing a-tubulin isoforms detectable by the terminal Tyr-specific antibody and the general a-tubulin antibody (Duckett and Lloyd, 1994;Wiesler et al, 2002;Wang et al, 2004). In this study, the Tyr antibody recognized Arabidopsis and tobacco tubulin from suspension cell cultures, resulting in a similar number and intensity of bands to those detected by the general a-tubulin antibody, whereas the Glu antibody did not (Fig.…”

“…This antibody has been used to detect tubulin acetylation, which varies considerably among cell and tissue types (Aström, 1992;Wang et al, 2004;Giannoutsou et al, 2012), at different developmental stages (Nakagawa et al, 2013), and as a result of hormonal treatments (Huang and Lloyd, 1999). Lys-40 is only partially conserved in plant a-tubulin isotypes.…”

Effects of post-translational modifications catalysed by pollen transglutaminase on the functional properties of microtubules and actin filaments

“…Tubulin is post-translationally modified to realize its multitude of functions during pollen tube growth [8]. On the other hand, post-translational modifications of actin are poorly known in eukaryotic cells [9].…”

Microtubule heterogeneity of <i>Ornithogalum umbellatum</i> ovary epidermal cells: non-stable cortical microtubules and stable lipotubuloid microtubules

“…Neuronal microtubules are the most differentiated; they play an important role in neuronal differentiation-regulation by controlling tubulin availability and modulating microtubule dy- [3,6]. In plants it has been observed that some PTMs of a tubulin preferentially appear in a tissue--specific manner [9].…”