Microtubule Minus-End Binding Protein CAMSAP2 and Kinesin-14 Motor KIFC3 Control Dendritic Microtubule Organization

Yujie Cao, Joanna Lipka, Riccardo Stucchi, Mithila Burute, Xingxiu Pan, Sybren Portegies, Roderick Tas, Jelmer Willems, Lena Will, Harold MacGillavry, Maarten Altelaar, Lukas C. Kapitein, Martin Harterink, Casper C. Hoogenraad

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Neuronal dendrites are characterized by an anti-parallel microtubule organization. The mechanism that regulates dendritic microtubule organization is still unclear. Cao et al. demonstrate that the microtubule minus-end binding protein CAMSAP2 and kinesin-14 motor KIFC3 work together to organize dendritic microtubules and control dendrite branching. Neuronal dendrites are characterized by an anti-parallel microtubule organization. The mixed oriented microtubules promote dendrite development and facilitate polarized cargo trafficking; however, the mechanism that regulates dendritic microtubule organization is still unclear. Here, we found that the kinesin-14 motor KIFC3 is important for organizing dendritic microtubules and to control dendrite development. The kinesin-14 motor proteins (Drosophila melanogaster Ncd, Saccharomyces cerevisiae Kar3, Saccharomyces pombe Pkl1, and Xenopus laevis XCTK2) are characterized by a C-terminal motor domain and are well described to organize the spindle microtubule during mitosis using an additional microtubule binding site in the N terminus [1–4]. In mammals, there are three kinesin-14 members, KIFC1, KIFC2, and KIFC3. It was recently shown that KIFC1 is important for organizing axonal microtubules in neurons, a process that depends on the two microtubule-interacting domains [5]. Unlike KIFC1, KIFC2 and KIFC3 lack the N-terminal microtubule binding domain and only have one microtubule-interacting domain, the motor domain [6, 7]. Thus, in order to regulate microtubule-microtubule crosslinking or sliding, KIFC2 and KIFC3 need to interact with additional microtubule binding proteins to connect two microtubules. We found that KIFC3 has a dendrite-specific distribution and interacts with microtubule minus-end binding protein CAMSAP2. Depletion of KIFC3 or CAMSAP2 results in increased microtubule dynamics during dendritic development. We propose a model in which CAMSAP2 anchors KIFC3 at microtubule minus ends and immobilizes microtubule arrays in dendrites.
Original languageEnglish
Pages (from-to)899-908.e6
JournalCurrent Biology
Volume30
Issue number5
DOIs
Publication statusPublished - 9 Mar 2020
Externally publishedYes

Funding

We thank Dr. Anna Akhmanova for sharing bio-mCherry and bio-GFP-N1 constructs. This work was supported by the Netherlands Organization for Scientific Research (NWO-ALW-VICI; 865.10.010; C.C.H.), the Netherlands Organization for Health Research and Development (ZonMW-TOP; 912.16.058; C.C.H.), the European Research Council (ERC) (ERC-consolidator; 617050; C.C.H.), and China Scholarship Council (CSC). Y.C. designed and performed the experiments, analyzed results, and wrote the manuscript; J.L. initiated the project and designed KIFC3 shRNAs; R.S. performed the mass spectrometry experiment; R.T. helped with super-resolution imaging; M.B. helped with the microtubule-severing experiment in COS7 and PEX imaging and quantification; L.W. and S.P. performed ex vivo electroporation and organotypic slice cultures; X.P. performed immunohistochemistry staining, imaging, and quantification for ex vivo experiments; J.W. helped with the CRISPR KI plasmid cloning; H.M. M.A. and L.C.K. gave valuable advice; and M.H. and C.C.H. designed the experiments, supervised the research, and wrote the manuscript. C.C.H. is an employee of Genentech, a member of the Roche group. The authors declare no additional competing interests. We thank Dr. Anna Akhmanova for sharing bio-mCherry and bio-GFP-N1 constructs. This work was supported by the Netherlands Organization for Scientific Research (NWO-ALW-VICI; 865.10.010 ; C.C.H.), the Netherlands Organization for Health Research and Development (ZonMW-TOP; 912.16.058 ; C.C.H.), the European Research Council (ERC) (ERC-consolidator; 617050 ; C.C.H.), and China Scholarship Council (CSC).

FundersFunder number
Nederlandse Organisatie voor Wetenschappelijk Onderzoek
China Scholarship Council
European Research Council
NWO-ALW-VICI865.10.010
Netherlands Organization for Health Research and Development912.16.058
Seventh Framework Programme617050

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