CEP78 functions downstream of CEP350 to control biogenesis of primary cilia by negatively regulating CP110 levels

André Brás Gonçalves; Sarah Kirstine Hasselbalch; Beinta Biskopstø Joensen; Sebastian Patzke; Pernille Martens; Signe Krogh Ohlsen; Mathieu Quinodoz; Konstantinos Nikopoulos; Reem Suleiman; Magnus Per Damsø Jeppesen; Catja Weiss; Søren Tvorup Christensen; Carlo Rivolta; Jens S. Andersen; Pietro Farinelli; Lotte Bang Pedersen

doi:10.7554/eLife.63731

CEP78 functions downstream of CEP350 to control biogenesis of primary cilia by negatively regulating CP110 levels

André Brás Gonçalves, Sarah Kirstine Hasselbalch, Beinta Biskopstø Joensen, Sebastian Patzke, Pernille Martens, Signe Krogh Ohlsen, Mathieu Quinodoz, Konstantinos Nikopoulos, Reem Suleiman, Magnus Per Damsø Jeppesen, Catja Weiss, Søren Tvorup Christensen, Carlo Rivolta, Jens S. Andersen, Pietro Farinelli^*, Lotte Bang Pedersen^*

^*Kontaktforfatter

Institut for Biokemi og Molekylær Biologi

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

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Abstract

CEP78 is a centrosomal protein implicated in ciliogenesis and ciliary length control, and mutations in the CEP78 gene cause retinal cone-rod dystrophy associated with hearing loss. However, the mechanism by which CEP78 affects cilia formation is unknown. Based on a recently discovered disease-causing CEP78 p.L150S mutation, we identified the disease-relevant interactome of CEP78. We confirmed that CEP78 interacts with the EDD1 -DYRK2-DDB1^VPRBP E3 ubiquitin lig- ase complex, which is involved in CP110 ubiquitination and degradation, and identified a novel interaction between CEP78 and CEP350 that is weakened by the CEP78^L150S mutation. We show that CEP350 promotes centrosomal recruitment and stability of CEP78, which in turn leads to cen- trosomal recruitment of EDD 1. Consistently, cells lacking CEP78 display significantly increased cellular and centrosomal levels of CP110, and depletion of CP110 in CEP78-deficient cells restored ciliation frequency to normal. We propose that CEP78 functions downstream of CEP350 to pro- mote ciliogenesis by negatively regulating CP110 levels via an EDD 1-dependent mechanism.

Originalsprog	Engelsk
Artikelnummer	e63731
Tidsskrift	eLife
Vol/bind	10
Antal sider	34
ISSN	2050-084X
DOI	https://doi.org/10.7554/eLife.63731
Status	Udgivet - aug. 2021

Bibliografisk note

Funding Information:
We thank Søren Lek Johansen and Maria Schrøder Holm for expert technical assistance, Benedicte Schultz Kappel and Frida Roikjer Rasmussen for help with plasmid generation, and Nynne Chris-tensen, Center for Advanced Bioimaging (CAB) Denmark, for help with 3D-SIM acquisition. Dr. Kay Schink kindly provided plasmids for lentivirus particle generation and assisted in 3D-SIM acquisition at OUH Radiumhospitalet. We are grateful to Drs. Peter K. Jackson, Tomoharu Kanie, Francesc Garcia-Gonzalo, Laurence Pelletier, Brian David Dynlacht, Anne-Marie Tassin, Maria Gavilan, Rosa M. Rios, Éric A. Cohen and Peter ten Dijke for reagents, and Dr. Michel Bornens for sharing original EM microcraphs. This study was supported grants from Independent Research Fund Denmark (#8020‐00162B to PF and LBP, and #8021-00425A to JSA), from the Carlsberg Foundation (#CF18‐0294 to LBP) and from the Swiss National Science Foundation (#176097 to CR).

Publisher Copyright:
© 2021, eLife Sciences Publications Ltd. All rights reserved.

Dokumenter og links

10.7554/eLife.63731Licens: CC BY

Open Access VersionForlagets udgivne version, 6,81 MBLicens: CC BY

Citationsformater

Gonçalves, A. B., Hasselbalch, S. K., Joensen, B. B., Patzke, S., Martens, P., Ohlsen, S. K., Quinodoz, M., Nikopoulos, K., Suleiman, R., Jeppesen, M. P. D., Weiss, C., Christensen, S. T., Rivolta, C., Andersen, J. S., Farinelli, P., & Pedersen, L. B. (2021). CEP78 functions downstream of CEP350 to control biogenesis of primary cilia by negatively regulating CP110 levels. eLife, 10, [e63731]. https://doi.org/10.7554/eLife.63731

@article{94bd4eae3adf4499a4e8ffea9e902f54,

title = "CEP78 functions downstream of CEP350 to control biogenesis of primary cilia by negatively regulating CP110 levels",

abstract = "CEP78 is a centrosomal protein implicated in ciliogenesis and ciliary length control, and mutations in the CEP78 gene cause retinal cone-rod dystrophy associated with hearing loss. However, the mechanism by which CEP78 affects cilia formation is unknown. Based on a recently discovered disease-causing CEP78 p.L150S mutation, we identified the disease-relevant interactome of CEP78. We confirmed that CEP78 interacts with the EDD1 -DYRK2-DDB1VPRBP E3 ubiquitin lig- ase complex, which is involved in CP110 ubiquitination and degradation, and identified a novel interaction between CEP78 and CEP350 that is weakened by the CEP78L150S mutation. We show that CEP350 promotes centrosomal recruitment and stability of CEP78, which in turn leads to cen- trosomal recruitment of EDD 1. Consistently, cells lacking CEP78 display significantly increased cellular and centrosomal levels of CP110, and depletion of CP110 in CEP78-deficient cells restored ciliation frequency to normal. We propose that CEP78 functions downstream of CEP350 to pro- mote ciliogenesis by negatively regulating CP110 levels via an EDD 1-dependent mechanism.",

author = "Gon{\c c}alves, {Andr{\'e} Br{\'a}s} and Hasselbalch, {Sarah Kirstine} and Joensen, {Beinta Biskopst{\o}} and Sebastian Patzke and Pernille Martens and Ohlsen, {Signe Krogh} and Mathieu Quinodoz and Konstantinos Nikopoulos and Reem Suleiman and Jeppesen, {Magnus Per Dams{\o}} and Catja Weiss and Christensen, {S{\o}ren Tvorup} and Carlo Rivolta and Andersen, {Jens S.} and Pietro Farinelli and Pedersen, {Lotte Bang}",

year = "2021",

month = aug,

doi = "10.7554/eLife.63731",

language = "English",

volume = "10",

journal = "eLife",

issn = "2050-084X",

publisher = "eLife Sciences Publications Ltd.",

}

Gonçalves, AB, Hasselbalch, SK, Joensen, BB, Patzke, S, Martens, P, Ohlsen, SK, Quinodoz, M, Nikopoulos, K, Suleiman, R, Jeppesen, MPD, Weiss, C, Christensen, ST, Rivolta, C, Andersen, JS, Farinelli, P & Pedersen, LB 2021, 'CEP78 functions downstream of CEP350 to control biogenesis of primary cilia by negatively regulating CP110 levels', eLife, bind 10, e63731. https://doi.org/10.7554/eLife.63731

TY - JOUR

T1 - CEP78 functions downstream of CEP350 to control biogenesis of primary cilia by negatively regulating CP110 levels

AU - Gonçalves, André Brás

AU - Hasselbalch, Sarah Kirstine

AU - Joensen, Beinta Biskopstø

AU - Patzke, Sebastian

AU - Martens, Pernille

AU - Ohlsen, Signe Krogh

AU - Quinodoz, Mathieu

AU - Nikopoulos, Konstantinos

AU - Suleiman, Reem

AU - Jeppesen, Magnus Per Damsø

AU - Weiss, Catja

AU - Christensen, Søren Tvorup

AU - Rivolta, Carlo

AU - Andersen, Jens S.

AU - Farinelli, Pietro

AU - Pedersen, Lotte Bang

PY - 2021/8

Y1 - 2021/8

N2 - CEP78 is a centrosomal protein implicated in ciliogenesis and ciliary length control, and mutations in the CEP78 gene cause retinal cone-rod dystrophy associated with hearing loss. However, the mechanism by which CEP78 affects cilia formation is unknown. Based on a recently discovered disease-causing CEP78 p.L150S mutation, we identified the disease-relevant interactome of CEP78. We confirmed that CEP78 interacts with the EDD1 -DYRK2-DDB1VPRBP E3 ubiquitin lig- ase complex, which is involved in CP110 ubiquitination and degradation, and identified a novel interaction between CEP78 and CEP350 that is weakened by the CEP78L150S mutation. We show that CEP350 promotes centrosomal recruitment and stability of CEP78, which in turn leads to cen- trosomal recruitment of EDD 1. Consistently, cells lacking CEP78 display significantly increased cellular and centrosomal levels of CP110, and depletion of CP110 in CEP78-deficient cells restored ciliation frequency to normal. We propose that CEP78 functions downstream of CEP350 to pro- mote ciliogenesis by negatively regulating CP110 levels via an EDD 1-dependent mechanism.

AB - CEP78 is a centrosomal protein implicated in ciliogenesis and ciliary length control, and mutations in the CEP78 gene cause retinal cone-rod dystrophy associated with hearing loss. However, the mechanism by which CEP78 affects cilia formation is unknown. Based on a recently discovered disease-causing CEP78 p.L150S mutation, we identified the disease-relevant interactome of CEP78. We confirmed that CEP78 interacts with the EDD1 -DYRK2-DDB1VPRBP E3 ubiquitin lig- ase complex, which is involved in CP110 ubiquitination and degradation, and identified a novel interaction between CEP78 and CEP350 that is weakened by the CEP78L150S mutation. We show that CEP350 promotes centrosomal recruitment and stability of CEP78, which in turn leads to cen- trosomal recruitment of EDD 1. Consistently, cells lacking CEP78 display significantly increased cellular and centrosomal levels of CP110, and depletion of CP110 in CEP78-deficient cells restored ciliation frequency to normal. We propose that CEP78 functions downstream of CEP350 to pro- mote ciliogenesis by negatively regulating CP110 levels via an EDD 1-dependent mechanism.

U2 - 10.7554/eLife.63731

DO - 10.7554/eLife.63731

M3 - Journal article

C2 - 34259627

AN - SCOPUS:85111833101

SN - 2050-084X

VL - 10

JO - eLife

JF - eLife

M1 - e63731

ER -

CEP78 functions downstream of CEP350 to control biogenesis of primary cilia by negatively regulating CP110 levels

Abstract

Bibliografisk note

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