NEDD1-S411 phosphorylation plays a critical function in the coordination of microtubule nucleation during mitosis

Author:

Timón Pérez Krystal1,Scrofani Jacopo1ORCID,Vernos Isabelle123ORCID

Affiliation:

1. Quantitative Cell Biology Program, Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology 1 , Barcelona 08003 , Spain

2. Universitat Pompeu Fabra (UPF) 2 , Barcelona 08003 , Spain

3. ICREA, Pg. Lluis Companys 23 3 , Barcelona 08010 , Spain

Abstract

ABSTRACT During mitosis, spindle assembly relies on centrosomal and acentrosomal microtubule nucleation pathways that all require the γ-Tubulin Ring Complex (γ-TuRC) and its adaptor protein NEDD1. The activity of these different pathways needs to be coordinated to ensure bipolar spindle assembly ( Cavazza et al., 2016) but the underlying mechanism is still unclear. Previous studies have identified three sites in NEDD1 (S377, S405 and S411) that when phosphorylated drive MT nucleation at the centrosomes, around the chromosomes and on pre-existing MTs respectively ( Lüders et al., 2006; Pinyol et al., 2013; Sdelci et al., 2012). Here we aimed at getting additional insights into the mechanism that coordinates the different MT nucleation pathways in dividing cells using a collection of HeLa stable inducible cell lines expressing NEDD1 phospho-variants at these three sites and Xenopus egg extracts. Our results provide further support for the essential role of phosphorylation at the three residues. Moreover, we directly demonstrate that S411 phosphorylation is essential for MT branching using TIRF microscopy in Xenopus egg extracts and we show that it plays a crucial role in ensuring the balance between centrosome and chromosome-dependent MT nucleation required for bipolar spindle assembly in mitotic cells.

Funder

Spanish Ministry of Economy, Industry and Competitiveness

Spanish Ministry of Economy and Competitiveness, ‘Centro de Excelencia Severo Ochoa’

CERCA Programme / Generalitat de Catalunya

Spanish Ministry of Economy and competitiveness

Ministry of Science, Innovation and Universities

Generalitat de Catalunya and intramural funds from the Centre for Genomic Regulation

CRG: Centre de Regulacio Genomica

Publisher

The Company of Biologists

Subject

General Agricultural and Biological Sciences,General Biochemistry, Genetics and Molecular Biology

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