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Δ3-tubulin impairs mitotic spindle morphology and increases nuclear size in pancreatic cancer cells

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Abstract

Cancer cell proliferation is affected by post-translational modifications of tubulin. Especially, overexpression or depletion of enzymes for modifications on the tubulin C-terminal region perturbs dynamic instability of the spindle body. Those modifications include processing of C-terminal amino acids of α-tubulin; detyrosination, and a removal of penultimate glutamic acid (Δ2). We previously found a further removal of the third last glutamic acid, which generates so-called Δ3-tubulin. The effects of Δ3-tubulin on spindle integrities and cell proliferation remain to be elucidated. In this study, we investigated the impacts of forced expression of Δ3-tubulin on the structure of spindle bodies and cell division in a pancreatic cancer cell line, PANC-1. Overexpression of HA-tagged Δ3-tubulin impaired the morphology and orientation of spindle bodies during cell division in PANC-1 cells. In particular, spindle bending was most significantly increased. Expression of EGFP-tagged Δ3-tubulin driven by the endogenous promoter of human TUBA1B also deformed and misoriented spindle bodies. Spindle bending and condensation defects were significantly observed by EGFP-Δ3-tubulin expression. Furthermore, EGFP-Δ3-tubulin expression increased the nuclear size in a dose-dependent manner of EGFP-Δ3-tubulin expression. The expression of EGFP-Δ3-tubulin tended to slow down cell proliferation. Taken together, our results demonstrate that Δ3-tubulin affects the spindle integrity and cell division.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

We thank Madoka Hamada and Yoko Hayashi for their technical assistance. This work was supported in part by JSPS Grants-in-Aids for Scientific Research (C) (22K08776) to K.U. and K.I., and by the Uehara Memorial Foundation (to K.I.). A part of this work was conducted in the Natural Science Center for Basic Research and Development at Hiroshima University (NBARD-00002) under the support from the MEXT Project to promote public utilization of advanced research infrastructure (Program for supporting the construction of core facilities; Grant Number JPMXS0441300023).

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Authors and Affiliations

  1. Department of Surgery, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8553, Japan

    Kenta Baba, Kenichiro Uemura & Shinya Takahashi

  2. Department of Anatomy and Developmental Biology, Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8553, Japan

    Kenta Baba, Ryota Nakazato, Faryal Ijaz & Koji Ikegami

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  1. Kenta Baba
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KB, FI, and KI designed the research; KB, FI, and RN carried out the experiments; KB, RN and KI analyzed the data; KU and ST organized the collaboration; and KB and KI wrote the paper. All authors reviewed the manuscript.

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Correspondence to Koji Ikegami.

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Baba, K., Uemura, K., Nakazato, R. et al. Δ3-tubulin impairs mitotic spindle morphology and increases nuclear size in pancreatic cancer cells. Med Mol Morphol 57, 59–67 (2024). https://doi.org/10.1007/s00795-023-00373-w

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