Two NEMO-like Ubiquitin-Binding Domains in CEP55 Differently Regulate Cytokinesis

Keïs Nabhane Said Halidi; Elisabeth Fontan; Alix Boucharlat; Laurianne Davignon; Marine Charpentier; Mikaël Boullé; Robert Weil; Alain Israël; Emmanuel Laplantine; Fabrice Agou

doi:10.1016/j.isci.2019.08.042

Two NEMO-like Ubiquitin-Binding Domains in CEP55 Differently Regulate Cytokinesis

iScience. 2019 Oct 25:20:292-309. doi: 10.1016/j.isci.2019.08.042. Epub 2019 Sep 25.

Affiliations

¹ Chemogenomic and Biological Screening Core Facility, C2RT, Departments of Cell Biology & Infection and of Structural Biology and Chemistry, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris CEDEX 15, France; Sorbonne Université, Collège doctoral, 75005 Paris, France.
² Chemogenomic and Biological Screening Core Facility, C2RT, Departments of Cell Biology & Infection and of Structural Biology and Chemistry, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris CEDEX 15, France.
³ Chemogenomic and Biological Screening Core Facility, C2RT, Departments of Cell Biology & Infection and of Structural Biology and Chemistry, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris CEDEX 15, France; Université de Paris, Sorbonne Paris Cité, Paris, France.
⁴ Signaling and Pathogenesis Laboratory, Department of Cell Biology & Infection, Institut Pasteur, C.N.R.S UMR 3691, 25 rue du Dr. Roux, 75724 Paris CEDEX 15, France.
⁵ Department of Cell Biology & Infection, Institut Pasteur, 75724 Paris CEDEX 15, France.
⁶ Signaling and Pathogenesis Laboratory, Department of Cell Biology & Infection, Institut Pasteur, C.N.R.S UMR 3691, 25 rue du Dr. Roux, 75724 Paris CEDEX 15, France. Electronic address: [email protected].
⁷ Chemogenomic and Biological Screening Core Facility, C2RT, Departments of Cell Biology & Infection and of Structural Biology and Chemistry, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris CEDEX 15, France. Electronic address: [email protected].

Abstract

CEP55 regulates the final critical step of cell division termed cytokinetic abscission. We report herein that CEP55 contains two NEMO-like ubiquitin-binding domains (UBDs), NOA and ZF, which regulate its function in a different manner. In vitro studies of isolated domains showed that NOA adopts a dimeric coiled-coil structure, whereas ZF is based on a UBZ scaffold. Strikingly, CEP55 knocked-down HeLa cells reconstituted with the full-length CEP55 ubiquitin-binding defective mutants, containing structure-guided mutations either in NOA^CEP55 or ZF^CEP55 domains, display severe abscission defects. In addition, the ZF^CEP55 can be functionally replaced by some ZF-based UBDs belonging to the UBZ family, indicating that the essential function of ZF^CEP55 is to act as ubiquitin receptor. Our work reveals an unexpected role of CEP55 in non-degradative ubiquitin signaling during cytokinetic abscission and provides a molecular basis as to how CEP55 mutations can lead to neurological disorders such as the MARCH syndrome.

Keywords: Biochemistry; Cell Biology; Protein Structure Aspects; Structural Biology.