Publication Date:
2022-10-27
Description:
© The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Hirst, W. G., Biswas, A., Mahalingan, K. K., & Reber, S. Differences in intrinsic tubulin dynamic properties contribute to spindle length control in Xenopus species. Current Biology, 30(11), (2020): 2184-2190.e5, doi: 10.1016/j.cub.2020.03.067.
Description:
The function of cellular organelles relates not only to their molecular composition but also to their size. However, how the size of dynamic mesoscale structures is established and maintained remains poorly understood [1, 2, 3]. Mitotic spindle length, for example, varies several-fold among cell types and among different organisms [4]. Although most studies on spindle size control focus on changes in proteins that regulate microtubule dynamics [5, 6, 7, 8], the contribution of the spindle’s main building block, the αβ-tubulin heterodimer, has yet to be studied. Apart from microtubule-associated proteins and motors, two factors have been shown to contribute to the heterogeneity of microtubule dynamics: tubulin isoform composition [9, 10] and post-translational modifications [11]. In the past, studying the contribution of tubulin and microtubules to spindle assembly has been limited by the fact that physiologically relevant tubulins were not available. Here, we show that tubulins purified from two closely related frogs, Xenopus laevis and Xenopus tropicalis, have surprisingly different microtubule dynamics in vitro. X. laevis microtubules combine very fast growth and infrequent catastrophes. In contrast, X. tropicalis microtubules grow slower and catastrophe more frequently. We show that spindle length and microtubule mass can be controlled by titrating the ratios of the tubulins from the two frog species. Furthermore, we combine our in vitro reconstitution assay and egg extract experiments with computational modeling to show that differences in intrinsic properties of different tubulins contribute to the control of microtubule mass and therefore set steady-state spindle length.
Description:
This article was prompted by our stay at the Marine Biological Laboratory (MBL), Woods Hole, MA in the summer of 2016 funded by the Princeton-Humboldt Strategic Partnership Grant together with the lab of Sabine Petry (Princeton University). We thank Jeff Woodruff (UT Southwestern), David Drechsel (IMP), and Marcus J. Taylor (MPI IB) for constructive criticism and comments on the manuscript and Helena Jambor for constructive comments on figure design. We thank the AMBIO imaging facility (Charité, Berlin) and Nikon at MBL for imaging support, Aliona Bogdanova and Barbara Borgonovo (MPI CBG) for their help with protein purification, and Francois Nedelec (University of Cambridge) for help with Cytosim. We are grateful to the Görlich lab (MPI BPC), in particular Bastian Hülsmann and Jens Krull, and the NXR for supply with X. tropicalis frogs. We thank Antonina Roll-Mecak (National Institute of Neurological Disorders and Stroke) for help with mass spectrometry analysis and discussions and Duck-Yeon Lee in the Biochemistry Core (National Heart, Lung and Blood Institute) for access to mass spectrometers. For mass spectrometry, we would like to acknowledge the assistance of Benno Kuropka and Chris Weise from the Core Facility BioSupraMol supported by the Deutsche Forschungsgemeinschaft (DFG). We thank all former and current members of the Reber lab for discussion and helpful advice, in particular, Christoph Hentschel and Soma Zsoter for technical assistance and Sebastian Reusch for help with tubulin purification. S.R. acknowledges funding from the IRI Life Sciences (Humboldt-Universität zu Berlin, Excellence Initiative/DFG). W.G.H. was supported by the Alliance Berlin Canberra co-funded by a grant from the Deutsche Forschungsgemeinschaft (DFG) for the International Research Training Group (IRTG) 2290 and the Australian National University. K.K.M. was supported by funds in the Roll-Mecak lab, intramural program of the National Institute of Neurological Disorders and Stroke.
Keywords:
Spindle scaling
;
Tubulin
;
Microtubule dynamics
;
Xenopus
;
Spindle length
Repository Name:
Woods Hole Open Access Server
Type:
Article
Permalink
