EB1 and EB3 regulate microtubule minus end organization and Golgi morphology

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.

• Publication year

  2017

• Venue

  Journal of Cell Biology

• Publication date

  2017-10-02

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1083/jcb.201701024PMID 28814570PMCID 5626540
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Automated Analysis of Intracellular Dynamic Processes.

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  2016influential reference
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  2016influential reference
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  2015cited by this paper
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  2014cited by this paper
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  2014cited by this paper
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  2014influential reference
• The centrosome–Golgi apparatus nexus

  2014cited by this paper
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  2014influential reference
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  2012influential reference
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