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      HSPB1 Facilitates the Formation of Non-Centrosomal Microtubules

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          Abstract

          The remodeling capacity of microtubules (MT) is essential for their proper function. In mammals, MTs are predominantly formed at the centrosome, but can also originate from non-centrosomal sites, a process that is still poorly understood. We here show that the small heat shock protein HSPB1 plays a role in the control of non-centrosomal MT formation. The HSPB1 expression level regulates the balance between centrosomal and non-centrosomal MTs. The HSPB1 protein can be detected specifically at sites of de novo forming non-centrosomal MTs, while it is absent from the centrosomes. In addition, we show that HSPB1 binds preferentially to the lattice of newly formed MTs in vitro, suggesting that its function occurs by stabilizing MT seeds. Our findings open new avenues for the understanding of the role of HSPB1 in the development, maintenance and protection of cells with specialized non-centrosomal MT arrays.

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          Most cited references48

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          NIH Image to ImageJ: 25 years of image analysis.

          For the past 25 years NIH Image and ImageJ software have been pioneers as open tools for the analysis of scientific images. We discuss the origins, challenges and solutions of these two programs, and how their history can serve to advise and inform other software projects.
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            Dynamics and mechanics of the microtubule plus end.

            An important function of microtubules is to move cellular structures such as chromosomes, mitotic spindles and other organelles around inside cells. This is achieved by attaching the ends of microtubules to cellular structures; as the microtubules grow and shrink, the structures are pushed or pulled around the cell. How do the ends of microtubules couple to cellular structures, and how does this coupling regulate the stability and distribution of the microtubules? It is now clear that there are at least three properties of a microtubule end: it has alternate structures; it has a biochemical transition defined by GTP hydrolysis; and it forms a distinct target for the binding of specific proteins. These different properties can be unified by thinking of the microtubule as a molecular machine, which switches between growing and shrinking modes. Each mode is associated with a specific end structure on which end-binding proteins can assemble to modulate dynamics and couple the dynamic properties of microtubules to the movement of cellular structures.
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              Asymmetric CLASP-dependent nucleation of noncentrosomal microtubules at the trans-Golgi network.

              Proper organization of microtubule arrays is essential for intracellular trafficking and cell motility. It is generally assumed that most if not all microtubules in vertebrate somatic cells are formed by the centrosome. Here we demonstrate that a large number of microtubules in untreated human cells originate from the Golgi apparatus in a centrosome-independent manner. Both centrosomal and Golgi-emanating microtubules need gamma-tubulin for nucleation. Additionally, formation of microtubules at the Golgi requires CLASPs, microtubule-binding proteins that selectively coat noncentrosomal microtubule seeds. We show that CLASPs are recruited to the trans-Golgi network (TGN) at the Golgi periphery by the TGN protein GCC185. In sharp contrast to radial centrosomal arrays, microtubules nucleated at the peripheral Golgi compartment are preferentially oriented toward the leading edge in motile cells. We propose that Golgi-emanating microtubules contribute to the asymmetric microtubule networks in polarized cells and support diverse processes including post-Golgi transport to the cell front.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2013
                24 June 2013
                : 8
                : 6
                : e66541
                Affiliations
                [1 ]Department of Molecular Genetics, VIB and University of Antwerp, Antwerpen, Belgium
                [2 ]Neurogenetics Laboratory, Institute Born Bunge, University of Antwerp, Antwerpen, Belgium
                [3 ]GROUP-ID Consortium, Laboratory for Immunoregulation and Mucosal Immunology, University of Ghent, Ghent, Belgium
                [4 ]Department of Molecular Biomedical Research, VIB, Ghent, Belgium
                Stanford University School of Medicine, United States of America
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: LAS BA VT SJ. Performed the experiments: LAS BA VDW SG. Analyzed the data: LAS BA SJ. Wrote the paper: LAS BA VT SJ.

                [¤]

                Current address: MRC Laboratory of Molecular Biology, Hills Road, Cambridge, United Kingdom

                Article
                PONE-D-13-07581
                10.1371/journal.pone.0066541
                3691211
                23826100
                bbfc4a3a-d0d9-4d32-8b6c-b580dfcd235b
                Copyright @ 2013

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 20 February 2013
                : 7 May 2013
                Page count
                Pages: 13
                Funding
                This work was supported in part by the special research fund and the Methusalem program of the University of Antwerp, the Fund for Scientific Research (FWO-Flanders), the Medical Foundation Queen Elisabeth (GSKE), the “Association Belge contre les Maladies Neuromusculaires” (ABMM), the American Muscular Dystrophy Association (MDA). We thank the Hercules foundation providing funding for the transmission electron and spinning disk microscopes. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology
                Biochemistry
                Cytochemistry
                Organelles
                Macromolecular Assemblies
                Biophysics
                Cell Motility
                Microtubules
                Genetics
                Gene Function
                Molecular Cell Biology
                Cellular Structures
                Cytoskeleton
                Cytometry
                Image Cytometry
                Neuroscience
                Neurobiology of Disease and Regeneration
                Medicine
                Neurology
                Motor Neuron Diseases

                Uncategorized
                Uncategorized

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