WDR47 protects neuronal microtubule minus ends from katanin-mediated severing

Robin R. Buijs; Jessica J.A. Hummel; Mithila Burute; Xingxiu Pan; Yujie Cao; Riccardo Stucchi; Maarten Altelaar; Anna Akhmanova; Lukas C. Kapitein; Casper C. Hoogenraad

doi:10.1016/j.celrep.2021.109371

WDR47 protects neuronal microtubule minus ends from katanin-mediated severing

Robin R. Buijs, Jessica J.A. Hummel, Mithila Burute, Xingxiu Pan, Yujie Cao, Riccardo Stucchi, Maarten Altelaar, Anna Akhmanova, Lukas C. Kapitein, Casper C. Hoogenraad^*

^*Corresponding author for this work

extern

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Axons and dendrites are long extensions of neurons that contain arrays of noncentrosomal microtubules. Calmodulin-regulated spectrin-associated proteins (CAMSAPs) bind to and stabilize free microtubule minus ends and are critical for proper neuronal development and function. Previous studies have shown that the microtubule-severing ATPase katanin interacts with CAMSAPs and limits the length of CAMSAP-decorated microtubule stretches. However, how CAMSAP and microtubule minus end dynamics are regulated in neurons is poorly understood. Here, we show that the neuron-enriched protein WDR47 interacts with CAMSAPs and is critical for axon and dendrite development. We find that WDR47 accumulates at CAMSAP2-decorated microtubules, is essential for maintaining CAMSAP2 stretches, and protects minus ends from katanin-mediated severing. We propose a model where WDR47 protects CAMSAP2 at microtubule minus ends from katanin activity to ensure proper stabilization of the neuronal microtubule network.

Original language	English
Article number	109371
Pages (from-to)	1-e7
Journal	Cell Reports
Volume	36
Issue number	2
DOIs	https://doi.org/10.1016/j.celrep.2021.109371
Publication status	Published - 13 Jul 2021

Bibliographical note

Funding Information:
We thank Prof. K. Jiang (Wuhan University, China) and Dr. G. Far?as (Utrecht University, the Netherlands) for sharing DNA constructs. This work was supported by the Netherlands Organization for Scientific Research (NWO-ALW-VICI to C.C.H. and NWO Graduate Program to R.R.B. [022.003.003]), the Netherlands Organization for Health Research and Development (ZonMW-TOP to C.C.H.) and the European Research Council (ERC) (ERC-consolidator to C.C.H.). R.R.B. designed and performed experiments, analyzed data, and wrote the manuscript. J.J.A.H. cloned constructs, performed pull-down experiments, and edited the manuscript. M.B. performed COS-7 experiments, analyzed data, and edited the manuscript. X.P. and Y.C. generated CRISPR KI constructs and performed experiments. R.S. performed the mass spectrometry experiment. M.A. supervised the mass spectrometry experiment. A.A. gave advice. L.C.K. supervised the COS-7 experiments. C.C.H. designed and supervised the research and wrote and edited the manuscript. C.C.H. is an employee of Genentech, Inc. a member of the Roche group.

Funding Information:
We thank Prof. K. Jiang (Wuhan University, China) and Dr. G. Farías (Utrecht University, the Netherlands) for sharing DNA constructs. This work was supported by the Netherlands Organization for Scientific Research ( NWO-ALW-VICI to C.C.H. and NWO Graduate Program to R.R.B. [022.003.003]), the Netherlands Organization for Health Research and Development ( ZonMW-TOP to C.C.H.) and the European Research Council (ERC) ( ERC-consolidator to C.C.H.).

Publisher Copyright:
© 2021 The Author(s)

Funding

We thank Prof. K. Jiang (Wuhan University, China) and Dr. G. Far?as (Utrecht University, the Netherlands) for sharing DNA constructs. This work was supported by the Netherlands Organization for Scientific Research (NWO-ALW-VICI to C.C.H. and NWO Graduate Program to R.R.B. [022.003.003]), the Netherlands Organization for Health Research and Development (ZonMW-TOP to C.C.H.) and the European Research Council (ERC) (ERC-consolidator to C.C.H.). R.R.B. designed and performed experiments, analyzed data, and wrote the manuscript. J.J.A.H. cloned constructs, performed pull-down experiments, and edited the manuscript. M.B. performed COS-7 experiments, analyzed data, and edited the manuscript. X.P. and Y.C. generated CRISPR KI constructs and performed experiments. R.S. performed the mass spectrometry experiment. M.A. supervised the mass spectrometry experiment. A.A. gave advice. L.C.K. supervised the COS-7 experiments. C.C.H. designed and supervised the research and wrote and edited the manuscript. C.C.H. is an employee of Genentech, Inc. a member of the Roche group. We thank Prof. K. Jiang (Wuhan University, China) and Dr. G. Farías (Utrecht University, the Netherlands) for sharing DNA constructs. This work was supported by the Netherlands Organization for Scientific Research ( NWO-ALW-VICI to C.C.H. and NWO Graduate Program to R.R.B. [022.003.003]), the Netherlands Organization for Health Research and Development ( ZonMW-TOP to C.C.H.) and the European Research Council (ERC) ( ERC-consolidator to C.C.H.).

Keywords

axon
CAMSAP
centrosome
cytoskeleton
dendrite
katanin
microtubule
neuron
WDR47

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title = "WDR47 protects neuronal microtubule minus ends from katanin-mediated severing",

abstract = "Axons and dendrites are long extensions of neurons that contain arrays of noncentrosomal microtubules. Calmodulin-regulated spectrin-associated proteins (CAMSAPs) bind to and stabilize free microtubule minus ends and are critical for proper neuronal development and function. Previous studies have shown that the microtubule-severing ATPase katanin interacts with CAMSAPs and limits the length of CAMSAP-decorated microtubule stretches. However, how CAMSAP and microtubule minus end dynamics are regulated in neurons is poorly understood. Here, we show that the neuron-enriched protein WDR47 interacts with CAMSAPs and is critical for axon and dendrite development. We find that WDR47 accumulates at CAMSAP2-decorated microtubules, is essential for maintaining CAMSAP2 stretches, and protects minus ends from katanin-mediated severing. We propose a model where WDR47 protects CAMSAP2 at microtubule minus ends from katanin activity to ensure proper stabilization of the neuronal microtubule network.",

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author = "Buijs, \{Robin R.\} and Hummel, \{Jessica J.A.\} and Mithila Burute and Xingxiu Pan and Yujie Cao and Riccardo Stucchi and Maarten Altelaar and Anna Akhmanova and Kapitein, \{Lukas C.\} and Hoogenraad, \{Casper C.\}",

note = "Funding Information: We thank Prof. K. Jiang (Wuhan University, China) and Dr. G. Far?as (Utrecht University, the Netherlands) for sharing DNA constructs. This work was supported by the Netherlands Organization for Scientific Research (NWO-ALW-VICI to C.C.H. and NWO Graduate Program to R.R.B. [022.003.003]), the Netherlands Organization for Health Research and Development (ZonMW-TOP to C.C.H.) and the European Research Council (ERC) (ERC-consolidator to C.C.H.). R.R.B. designed and performed experiments, analyzed data, and wrote the manuscript. J.J.A.H. cloned constructs, performed pull-down experiments, and edited the manuscript. M.B. performed COS-7 experiments, analyzed data, and edited the manuscript. X.P. and Y.C. generated CRISPR KI constructs and performed experiments. R.S. performed the mass spectrometry experiment. M.A. supervised the mass spectrometry experiment. A.A. gave advice. L.C.K. supervised the COS-7 experiments. C.C.H. designed and supervised the research and wrote and edited the manuscript. C.C.H. is an employee of Genentech, Inc. a member of the Roche group. Funding Information: We thank Prof. K. Jiang (Wuhan University, China) and Dr. G. Far{\'i}as (Utrecht University, the Netherlands) for sharing DNA constructs. This work was supported by the Netherlands Organization for Scientific Research ( NWO-ALW-VICI to C.C.H. and NWO Graduate Program to R.R.B. [022.003.003]), the Netherlands Organization for Health Research and Development ( ZonMW-TOP to C.C.H.) and the European Research Council (ERC) ( ERC-consolidator to C.C.H.). Publisher Copyright: {\textcopyright} 2021 The Author(s)",

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doi = "10.1016/j.celrep.2021.109371",

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T1 - WDR47 protects neuronal microtubule minus ends from katanin-mediated severing

AU - Buijs, Robin R.

AU - Hummel, Jessica J.A.

AU - Burute, Mithila

AU - Pan, Xingxiu

AU - Cao, Yujie

AU - Stucchi, Riccardo

AU - Altelaar, Maarten

AU - Akhmanova, Anna

AU - Kapitein, Lukas C.

AU - Hoogenraad, Casper C.

N1 - Funding Information: We thank Prof. K. Jiang (Wuhan University, China) and Dr. G. Far?as (Utrecht University, the Netherlands) for sharing DNA constructs. This work was supported by the Netherlands Organization for Scientific Research (NWO-ALW-VICI to C.C.H. and NWO Graduate Program to R.R.B. [022.003.003]), the Netherlands Organization for Health Research and Development (ZonMW-TOP to C.C.H.) and the European Research Council (ERC) (ERC-consolidator to C.C.H.). R.R.B. designed and performed experiments, analyzed data, and wrote the manuscript. J.J.A.H. cloned constructs, performed pull-down experiments, and edited the manuscript. M.B. performed COS-7 experiments, analyzed data, and edited the manuscript. X.P. and Y.C. generated CRISPR KI constructs and performed experiments. R.S. performed the mass spectrometry experiment. M.A. supervised the mass spectrometry experiment. A.A. gave advice. L.C.K. supervised the COS-7 experiments. C.C.H. designed and supervised the research and wrote and edited the manuscript. C.C.H. is an employee of Genentech, Inc. a member of the Roche group. Funding Information: We thank Prof. K. Jiang (Wuhan University, China) and Dr. G. Farías (Utrecht University, the Netherlands) for sharing DNA constructs. This work was supported by the Netherlands Organization for Scientific Research ( NWO-ALW-VICI to C.C.H. and NWO Graduate Program to R.R.B. [022.003.003]), the Netherlands Organization for Health Research and Development ( ZonMW-TOP to C.C.H.) and the European Research Council (ERC) ( ERC-consolidator to C.C.H.). Publisher Copyright: © 2021 The Author(s)

PY - 2021/7/13

Y1 - 2021/7/13

N2 - Axons and dendrites are long extensions of neurons that contain arrays of noncentrosomal microtubules. Calmodulin-regulated spectrin-associated proteins (CAMSAPs) bind to and stabilize free microtubule minus ends and are critical for proper neuronal development and function. Previous studies have shown that the microtubule-severing ATPase katanin interacts with CAMSAPs and limits the length of CAMSAP-decorated microtubule stretches. However, how CAMSAP and microtubule minus end dynamics are regulated in neurons is poorly understood. Here, we show that the neuron-enriched protein WDR47 interacts with CAMSAPs and is critical for axon and dendrite development. We find that WDR47 accumulates at CAMSAP2-decorated microtubules, is essential for maintaining CAMSAP2 stretches, and protects minus ends from katanin-mediated severing. We propose a model where WDR47 protects CAMSAP2 at microtubule minus ends from katanin activity to ensure proper stabilization of the neuronal microtubule network.

AB - Axons and dendrites are long extensions of neurons that contain arrays of noncentrosomal microtubules. Calmodulin-regulated spectrin-associated proteins (CAMSAPs) bind to and stabilize free microtubule minus ends and are critical for proper neuronal development and function. Previous studies have shown that the microtubule-severing ATPase katanin interacts with CAMSAPs and limits the length of CAMSAP-decorated microtubule stretches. However, how CAMSAP and microtubule minus end dynamics are regulated in neurons is poorly understood. Here, we show that the neuron-enriched protein WDR47 interacts with CAMSAPs and is critical for axon and dendrite development. We find that WDR47 accumulates at CAMSAP2-decorated microtubules, is essential for maintaining CAMSAP2 stretches, and protects minus ends from katanin-mediated severing. We propose a model where WDR47 protects CAMSAP2 at microtubule minus ends from katanin activity to ensure proper stabilization of the neuronal microtubule network.

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KW - CAMSAP

KW - centrosome

KW - cytoskeleton

KW - dendrite

KW - katanin

KW - microtubule

KW - neuron

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ER -

WDR47 protects neuronal microtubule minus ends from katanin-mediated severing

Abstract

Bibliographical note

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Keywords

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