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p53 is associated with cellular microtubules and is transported to the nucleus by dynein

Nature Cell Biology volume 2, pages 709–717 (2000)Cite this article

Abstract

Here we show that p53 protein is physically associated with tubulin in vivo and in vitro, and that it localizes to cellular microtubules. Treatment with vincristine or paclitaxel before DNA-damage or before leptomycin B treatment reduces nuclear accumulation of p53 and expression of mdm2 and p21. Overexpression of dynamitin or microinjection of anti-dynein antibody before DNA damage abrogates nuclear accumulation of p53. Our results indicate that transport of p53 along microtubules is dynein-dependent. The first 25 amino acids of p53 contain the residues that are essential for binding to microtubules. We propose that functional microtubules and the dynein motor protein participate in transport of p53 and facilitate its accumulation in the nucleus after DNA damage.

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Figure 1: p53 co-immunoprecipitates with tubulin.
Figure 2: Wild-type and mutant p53 bind preferentially to polymerized tubulin.
Figure 3: Co-localization of p53 with cellular microtubules.
Figure 4: Microtubule-perturbing drugs impair nuclear accumulation of p53 and induction of mdm2 and p21 after DNA damage.
Figure 5: Pretreatment with paclitaxel or vincristine abrogates the effect of Leptomycin B on p53.
Figure 6: The microtubule–motor complex dynein/dynactin is involved in transport of p53.
Figure 7: Purified p53 binds to purified microtubules in vitro.
Figure 8: The N terminus of p53 is important for microtubule binding.

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Acknowledgements

We thank H. Gray and U. Roongta for help with expression and purification of His–p53 fusion protein. We are grateful to M. Clarke and C. Harris for providing full-length wild-type p53 and p53 (Δ342–end) plasmids, respectively. We also thank R. Vallee for providing the dynamitin plasmid, and L. Neckers for providing Leptomycin B. We are indebted to R. Klausner for insightful discussion.

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Authors and Affiliations

  1. Division of Clinical Sciences, Medicine Branch, National Cancer Institute, National Institutes of Health, 9000 Rockville Pike, Bethesda, 20892, Maryland, USA

    Paraskevi Giannakakou, Yvona Ward, Mikhail V. Blagosklonny & Tito Fojo

  2. Laboratory of Integrative and Medical Biophysics, National Institute of Child Health and Human Development, National Institutes of Health , 9000 Rockville Pike, Bethesda, 20892, Maryland , USA

    Dan L. Sackett

  3. Department of Oncology Drug Discovery, Bristol–Myers Squibb Pharmaceutical Research Institute, PO Box 4000 , Princeton, 08543-4000, New Jersey, USA

    Kevin R. Webster

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  1. Paraskevi Giannakakou
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Correspondence to Paraskevi Giannakakou.

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Giannakakou, P., Sackett, D., Ward, Y. et al. p53 is associated with cellular microtubules and is transported to the nucleus by dynein. Nat Cell Biol 2, 709–717 (2000). https://doi.org/10.1038/35036335

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