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      Positioning centrioles and centrosomes

      review-article
      1 , , 1 ,
      The Journal of Cell Biology
      Rockefeller University Press

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          Abstract

          Hannaford and Rusan summarize the direct and indirect transport mechanisms by which centrosomes and centrioles are positioned in cells.

          Abstract

          Centrosomes are the primary microtubule organizer in eukaryotic cells. In addition to shaping the intracellular microtubule network and the mitotic spindle, centrosomes are responsible for positioning cilia and flagella. To fulfill these diverse functions, centrosomes must be properly located within cells, which requires that they undergo intracellular transport. Importantly, centrosome mispositioning has been linked to ciliopathies, cancer, and infertility. The mechanisms by which centrosomes migrate are diverse and context dependent. In many cells, centrosomes move via indirect motor transport, whereby centrosomal microtubules engage anchored motor proteins that exert forces on those microtubules, resulting in centrosome movement. However, in some cases, centrosomes move via direct motor transport, whereby the centrosome or centriole functions as cargo that directly binds molecular motors which then walk on stationary microtubules. In this review, we summarize the mechanisms of centrosome motility and the consequences of centrosome mispositioning and identify key questions that remain to be addressed.

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          Genes and molecular pathways underpinning ciliopathies

          Motile and non-motile primary cilia are nearly ubiquitous cellular organelles. Dysfunction of cilia is being found to cause increasing numbers of diseases that are known as ciliopathies. The characterization of ciliopathy-associated proteins and phenotypes is increasing our understanding of how cilia are formed and compartmentalized and how they function to maintain human health.
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            The bipolar mitotic kinesin Eg5 moves on both microtubules that it crosslinks.

            During cell division, mitotic spindles are assembled by microtubule-based motor proteins. The bipolar organization of spindles is essential for proper segregation of chromosomes, and requires plus-end-directed homotetrameric motor proteins of the widely conserved kinesin-5 (BimC) family. Hypotheses for bipolar spindle formation include the 'push-pull mitotic muscle' model, in which kinesin-5 and opposing motor proteins act between overlapping microtubules. However, the precise roles of kinesin-5 during this process are unknown. Here we show that the vertebrate kinesin-5 Eg5 drives the sliding of microtubules depending on their relative orientation. We found in controlled in vitro assays that Eg5 has the remarkable capability of simultaneously moving at approximately 20 nm s(-1) towards the plus-ends of each of the two microtubules it crosslinks. For anti-parallel microtubules, this results in relative sliding at approximately 40 nm s(-1), comparable to spindle pole separation rates in vivo. Furthermore, we found that Eg5 can tether microtubule plus-ends, suggesting an additional microtubule-binding mode for Eg5. Our results demonstrate how members of the kinesin-5 family are likely to function in mitosis, pushing apart interpolar microtubules as well as recruiting microtubules into bundles that are subsequently polarized by relative sliding.
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              Phosphorylation by p34cdc2 regulates spindle association of human Eg5, a kinesin-related motor essential for bipolar spindle formation in vivo.

              We have isolated a human homolog of Xenopus Eg5, a kinesin-related motor protein implicated in the assembly and dynamics of the mitotic spindle. We report that microinjection of antibodies against human Eg5 (HsEg5) blocks centrosome migration and causes HeLa cells to arrest in mitosis with monoastral microtubule arrays. Furthermore, an evolutionarily conserved cdc2 phosphorylation site (Thr-927) in HsEg5 is phosphorylated specifically during mitosis in HeLa cells and by p34cdc2/cyclin B in vitro. Mutation of Thr-927 to nonphosphorylatable residues prevents HsEg5 from binding to centrosomes, indicating that phosphorylation controls the association of this motor with the spindle apparatus. These results indicate that HsEg5 is required for establishing a bipolar spindle and that p34cdc2 protein kinase directly regulates its localization.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Writing - original draftRole: Writing - review & editing
                Role: ConceptualizationRole: ResourcesRole: SupervisionRole: Writing - review & editing
                Journal
                J Cell Biol
                J Cell Biol
                jcb
                The Journal of Cell Biology
                Rockefeller University Press
                0021-9525
                1540-8140
                01 April 2024
                21 March 2024
                21 March 2024
                : 223
                : 4
                : e202311140
                Affiliations
                [1 ]Cell and Developmental Biology Center, National Heart, Lung, and Blood Institute, National Institutes of Health; , Bethesda, MD, USA
                Author notes
                Correspondence to Matthew R. Hannaford: matthew.hannaford@ 123456nih.gov
                Nasser M. Rusan: nasser@ 123456nih.gov

                Disclosures: The authors declare no competing interests exist.

                Author information
                https://orcid.org/0000-0001-7772-0450
                https://orcid.org/0000-0002-4194-1072
                Article
                jcb.202311140
                10.1083/jcb.202311140
                10959756
                38512059
                5bae3ec1-6e88-4818-af25-00f9f608747f
                This is a work of the U.S. Government and is not subject to copyright protection in the United States. Foreign copyrights may apply.

                This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

                History
                : 14 December 2023
                : 23 February 2024
                : 26 February 2024
                Funding
                Funded by: Division of Intramural Research, DOI http://dx.doi.org/10.13039/100006492;
                Funded by: National Heart, Lung, and Blood Institute, DOI http://dx.doi.org/10.13039/100000050;
                Award ID: 1ZIAHL006126
                Funded by: National Institutes of Health, DOI http://dx.doi.org/10.13039/100000002;
                Categories
                Review
                Development
                Cytoskeleton
                Cell Cycle and Division

                Cell biology
                Cell biology

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