Microtubule minus-end stabilization by polymerization-driven CAMSAP deposition.
Microtubules are cytoskeletal polymers with two structurally and functionally distinct ends, the plus- and the minus-end. Here, we focus on the mechanisms underlying the regulation of microtubule minus-ends by the CAMSAP/Nezha/Patronin protein family. We show that CAMSAP2 is required for the proper organization and stabilization of interphase microtubules and directional cell migration. By combining live-cell imaging and in vitro reconstitution of microtubule assembly from purified components with laser microsurgery, we demonstrate that CAMSAPs regulate microtubule minus-end growth and are specifically deposited on the lattice formed by microtubule minus-end polymerization. This process leads to the formation of CAMSAP-decorated microtubule stretches, which are stabilized from both ends and serve as sites of noncentrosomal microtubule outgrowth. The length of the stretches is regulated by the microtubule-severing protein katanin, which interacts with CAMSAPs. Our data thus indicate that microtubule minus-end assembly drives the stabilization of noncentrosomal microtubules and that katanin regulates this process.
Cell Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 Utrecht, the Netherlands.
Kai
K
Jiang
Cell Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 Utrecht, the Netherlands.
Shasha
S
Hua
Cell Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 Utrecht, the Netherlands.
Renu
R
Mohan
Cell Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 Utrecht, the Netherlands.
Ilya
I
Grigoriev
Cell Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 Utrecht, the Netherlands.
Kah Wai
KW
Yau
Cell Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 Utrecht, the Netherlands; Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research, Utrecht Institute for Pharmaceutical Sciences and The Netherlands Proteomics Centre, Utrecht University, Padualaan 8, 3584 Utrecht, the Netherlands.
Qingyang
Q
Liu
Cell Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 Utrecht, the Netherlands.
Eugene A
EA
Katrukha
Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research, Utrecht Institute for Pharmaceutical Sciences and The Netherlands Proteomics Centre, Utrecht University, Padualaan 8, 3584 Utrecht, the Netherlands.
A F Maarten
AF
Altelaar
Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research, Utrecht Institute for Pharmaceutical Sciences and The Netherlands Proteomics Centre, Utrecht University, Padualaan 8, 3584 Utrecht, the Netherlands.
Albert J R
AJ
Heck
Cell Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 Utrecht, the Netherlands.
Casper C
CC
Hoogenraad
Cell Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 Utrecht, the Netherlands. Electronic address: a.akhmanova@uu.nl.
Anna
A
Akhmanova
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D018385:Centrosome / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D000074201:Katanin; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*
C573192:CAMSAP2 protein, human; D003598:Cytoskeletal Proteins; D008869:Microtubule-Associated Proteins; D000251:Adenosine Triphosphatases; D000074201:Katanin