Structure of the RZZ complex and molecular basis of its interaction with Spindly

Mosalaganti, Shyamal; Keller, Jenny; Altenfeld, Anika; Winzker, Michael; Rombaut, Pascaline; Saur, Michael; Petrovic, Arsen; Wehenkel, Annemarie; Wohlgemuth, Sabine; Müller, Franziska; Maffini, Stefano; Bange, Tanja; Herzog, Franz ; Waldmann, Herbert; Raunser, Stefan and Musacchio, Andrea (2017): Structure of the RZZ complex and molecular basis of its interaction with Spindly. In: Journal of cell biology, Vol. 216, No. 4: pp. 961-981

Abstract

Kinetochores are macromolecular assemblies that connect chromosomes to spindle microtubules (MTs) during mitosis. The metazoan-specific ≈800-kD ROD-Zwilch-ZW10 (RZZ) complex builds a fibrous corona that assembles on mitotic kinetochores before MT attachment to promote chromosome alignment and robust spindle assembly checkpoint signaling. In this study, we combine biochemical reconstitutions, single-particle electron cryomicroscopy, cross-linking mass spectrometry, and structural modeling to build a complete model of human RZZ. We find that RZZ is structurally related to self-assembling cytosolic coat scaffolds that mediate membrane cargo trafficking, including Clathrin, Sec13-Sec31, and αβ'ε-COP. We show that Spindly, a dynein adaptor, is related to BicD2 and binds RZZ directly in a farnesylation-dependent but membrane-independent manner. Through a targeted chemical biology approach, we identify ROD as the Spindly farnesyl receptor. Our results suggest that RZZ is dynein's cargo at human kinetochores.

• Item Type: Journal article
• EU Funded Grant Agreement Number: 638218
• EU Projects: Horizon 2020 > ERC Grants > ERC Starting Grant > ERC Grant 638218: MolStruKT - Molecular structure and cell cycle regulated assembly of the kinetochore
• Faculties: Chemistry and Pharmacy > Department of Biochemistry; Chemistry and Pharmacy > GeneCenter
• Subjects: 500 Science > 540 Chemistry
• Language: English