Post-translational modifications and multiple tubulin isoforms in Nicotiana tabacum L. cells |
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Authors: | Andrey Smertenko Yaroslav Blume Vladimír Viklický Zdeněk Opatrný Pavel Dráber |
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Institution: | (1) Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20 Praha 4, Czech Republic ,;(2) Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of the Ukraine, Zabolotnogo str. 148, GSP-22, 252022 Kiev, Ukraine , UA;(3) Institute of Crop Production, Drnovská 507, 161 06, Praha 6, Czech Republic, |
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Abstract: | Distribution of post-translationally modified tubulins in cells of Nicotiana tabacum L. was analysed using a panel of specific antibodies. Polyglutamylated, tyrosinated, nontyrosinated, acetylated and Δ2-tubulin
variants were detected on α-tubulin subunits; polyglutamylation was also found on β-tubulin subunits. Modified tubulins were
detected by immunofluorescence microscopy in interphase microtubules, preprophase bands, mitotic spindles as well as in phragmoplasts.
They were, however, located differently in the various microtubule structures. The antibodies against tyrosinated, acetylated
and polyglutamylated tubulins gave uniform staining along all microtubules, while antibodies against nontyrosinated and Δ2-tubulin
provided dot-like staining of interphase microtubules. Additionally, immunoreactivity of antibodies against acetylated and
Δ2-tubulins was strong in the pole regions of mitotic spindles. High-resolution isoelectric focusing revealed 22 tubulin charge
variants in N. tabacum suspension cells. Immunoblotting with antibodies TU-01 and TU-06 against conserved antigenic determinants of α- and β-tubulin
molecules, respectively, revealed that 11 isoforms belonged to the α-subunit and 11 isoforms to the β-subunit. Whereas antibodies
against polyglutamylated, tyrosinated and acetylated tubulins reacted with several α-tubulin isoforms, antibodies against
nontyrosinated and Δ2-tubulin reacted with only one. The combined data demonstrate that plant tubulin is extensively post-translationally
modified and that these modifications participate in the generation of plant tubulin polymorphism.
Received: 2 May 1996 / Accepted: 16 September 1996 |
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Keywords: | : High-resolution isoelectric focusing Immuno fluorescence microscopy Nicotiana microtubules Post-translational modifications of tubulin Tubulin isoforms |
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