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Post-translational modifications and multiple tubulin isoforms in Nicotiana tabacum L. cells

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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 dotlike 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 posttranslationally modified and that these modifications participate in the generation of plant tubulin polymorphism.

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Abbreviations

HRIF:

high-resolution isoelectric focusing

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Authors and Affiliations

  1. Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20, Praha 4, Czech Republic

    Andrey Smertenko, Vladimír Viklický & Pavel Dráber

  2. Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of the Ukraine, Zabolotnogo str. 148, GSP-22, 252022, Kiev, Ukraine

    Andrey Smertenko & Yaroslav Blume

  3. Institute of Crop Production, Drnovská 507, 161 06, Praha 6, Czech Republic

    Zdeněk Opatrný

Authors
  1. Andrey Smertenko
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  2. Yaroslav Blume
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  3. Vladimír Viklický
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  4. Zdeněk Opatrný
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  5. Pavel Dráber
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Correspondence to Pavel Dráber.

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Smertenko, A., Blume, Y., Viklický, V. et al. Post-translational modifications and multiple tubulin isoforms in Nicotiana tabacum L. cells. Planta 201, 349–358 (1997). https://doi.org/10.1007/s004250050077

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  • DOI: https://doi.org/10.1007/s004250050077

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