EB1 and EB3 regulate microtubule minus end organization and Golgi morphology

• Published in: The Journal of cell biology Vol. 216; no. 10; pp. 3179 - 3198
• Main Authors: Yang, Chao, Wu, Jingchao, de Heus, Cecilia, Grigoriev, Ilya, Liv, Nalan, Yao, Yao, Smal, Ihor, Meijering, Erik, Klumperman, Judith, Qi, Robert Z, Akhmanova, Anna
• Format: Journal Article
• Language: English
• Published: United States Rockefeller University Press 02-10-2017; The Rockefeller University Press
• Subjects: Binding; Biotechnology; Cell division; Cell lines; Cell migration; Cell Movement - physiology; Cell Polarity - physiology; Clustering; CRISPR; Cytoplasm; Cytoskeleton; Focal Adhesions - genetics; Focal Adhesions - metabolism; Golgi apparatus; Golgi Apparatus - genetics; Golgi Apparatus - metabolism; HeLa Cells; Humans; Interphase - physiology; Intracellular Membranes - metabolism; Membranes; Microtubule-Associated Proteins - genetics; Microtubule-Associated Proteins - metabolism; Microtubules; Microtubules - genetics; Microtubules - metabolism; Morphology; Muscle Proteins - genetics; Muscle Proteins - metabolism; Nuclear Proteins - genetics; Nuclear Proteins - metabolism; Polarity; Polymerization; Proteins
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.201701024

End-binding proteins (EBs) are the core components of microtubule plus end tracking protein complexes, but it is currently unknown whether they are essential for mammalian microtubule organization. Here, by using CRISPR/Cas9-mediated knockout technology, we generated stable cell lines lacking EB2 and EB3 and the C-terminal partner-binding half of EB1. These cell lines show only mild defects in cell division and microtubule polymerization. However, the length of CAMSAP2-decorated stretches at noncentrosomal microtubule minus ends in these cells is reduced, microtubules are detached from Golgi membranes, and the Golgi complex is more compact. Coorganization of microtubules and Golgi membranes depends on the EB1/EB3-myomegalin complex, which acts as membrane-microtubule tether and counteracts tight clustering of individual Golgi stacks. Disruption of EB1 and EB3 also perturbs cell migration, polarity, and the distribution of focal adhesions. EB1 and EB3 thus affect multiple interphase processes and have a major impact on microtubule minus end organization.