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The state of the guanosine nucleotide allosterically affects the interfaces of tubulin in protofilament

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Abstract

The dynamics of microtubules is essential for many microtubule-dependent cellular functions such as the mitosis. It has been recognized for a long time that GTP hydrolysis in αβ-tubulin polymers plays a critical role in this dynamics. However, the effects of the changes in the nature of the guanosine nucleotide at the E-site in β-tubulin on microtubule structure and stability are still not well understood. In the present work, we performed all-atom molecular dynamics simulations of a αβα-tubulin heterotrimer harboring a guanosine nucleotide in three different states at the E-site: GTP, GDP-Pi and GDP. We found that changes in the nucleotide state is associated with significant conformational variations at the α-tubulin N- and β-tubulin M-loops which impact the interactions between tubulin protofilaments. The results also show that GTP hydrolysis reduces αβ-tubulin interdimer contacts in favor of intradimer interface. From an atomistic point view, we propose a role for α-tubulin glutamate residue 254 in catalytic magnesium coordination and identified a water molecule in the nucleotide binding pocket which is most probably required for nucleotide hydrolysis. Finally, the results are discussed with reference to the role of taxol in microtubule stability and the recent tubulin-sT2R crystal structures.

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Abbreviations

GTP:

Guanosine triphosphate

GDP:

Guanosine diphosphate

Pi:

Inorganic phosphate

MT:

Microtubule

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Author information

Author notes

  1. Philippe Manivet

    Present address: Institut National de la Santé et de la Recherche Médicale (INSERM), UMRS 942, Hôpital Lariboisière, Evry, France

Authors and Affiliations

  1. Institut National de la Santé et de la Recherche Médicale (INSERM), UMRS 829, Evry, France

    Joseph R. André, Marie-Jeanne Clément, Elisabeth Adjadj, Flavio Toma & Patrick A. Curmi

  2. Département de Biologie, IBGBI, Université d’Evry-Val-d’Essonne, Evry, France

    Philippe Manivet

  3. APHP, GHU Saint Louis-Lariboisière-F Widal, Pole B2P, Hôpital Lariboisière, service de Biochimie et de Biologie moléculaire, Unité de Biologie Clinique Structurale, 2 rue Ambroise Paré, 75475, Paris cedex 10, France

    Philippe Manivet

  4. Université Evry-Val d’Essonne, Evry, France

    Joseph R. André, Marie-Jeanne Clément, Elisabeth Adjadj, Flavio Toma & Patrick A. Curmi

  5. Laboratoire Structure-Activité des Biomolécules Normales et Pathologiques, Evry, France

    Joseph R. André, Marie-Jeanne Clément, Elisabeth Adjadj, Flavio Toma & Patrick A. Curmi

  6. BioQuanta Corporation, Aurora, CO, USA

    Joseph R. André

  7. Institut National de la Santé et de la Recherche Médicale (INSERM), UMRS 829, SABNP, Evry, France

    Philippe Manivet

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  1. Joseph R. André
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Correspondence to Philippe Manivet.

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André, J.R., Clément, MJ., Adjadj, E. et al. The state of the guanosine nucleotide allosterically affects the interfaces of tubulin in protofilament. J Comput Aided Mol Des 26, 397–407 (2012). https://doi.org/10.1007/s10822-012-9566-x

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