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Uwe Knippschild - One of the best experts on this subject based on the ideXlab platform.
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interaction of casein kinase 1 delta ck1delta with post golgi structures microtubules and the Spindle Apparatus
European Journal of Cell Biology, 2000Co-Authors: Lars Behrend, Jochen Heukeshoven, Gabriel Rutter, Wolfgang Deppert, Marion Kurth, Martin Stoter, Uwe KnippschildAbstract:Summary Members of the casein kinase 1 family of serine/threonine kinases are highly conserved from yeast to mammals and seem to play an important role in vesicular trafficking, DNA repair, cell cycle progression and cytokinesis. We here report that in interphase cells of various mammalian species casein kinase 1 delta (CK1δ) specifically interacts with the trans Golgi network and cytoplasmic, granular particles that associate with microtubules. Furthermore, at mitosis CK1δ is recruited to the Spindle Apparatus and the centrosomes in cells, which have been exposed to DNA-damaging agents like etoposide or γ-irradiation. In addition, determination of the affinity of CK1δ to different tubulin isoforms in immunoprecipitation-Western analysis revealed a dramatically enhanced complex formation between CK1δ and tubulins from mitotic extracts after introducing DNA damage. The high affinity of CK1δ to the Spindle Apparatus in DNA-damaged cells and its ability to phosphorylate several microtubule-associated proteins points to a regulatory role of CK1δ at mitosis.
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interaction of casein kinase 1 delta ck1delta with post golgi structures microtubules and the Spindle Apparatus
European Journal of Cell Biology, 2000Co-Authors: Lars Behrend, Jochen Heukeshoven, Gabriel Rutter, Wolfgang Deppert, Marion Kurth, Martin Stoter, Uwe KnippschildAbstract:Members of the casein kinase 1 family of serine/threonine kinases are highly conserved from yeast to mammals and seem to play an important role in vesicular trafficking, DNA repair, cell cycle progression and cytokinesis. We here report that in interphase cells of various mammalian species casein kinase 1 delta (CK1delta) specifically interacts with the trans Golgi network and cytoplasmic, granular particles that associate with microtubules. Furthermore, at mitosis CK1delta is recruited to the Spindle Apparatus and the centrosomes in cells, which have been exposed to DNA-damaging agents like etoposide or gammairradiation. In addition, determination of the affinity of CK1delta to different tubulin isoforms in immunoprecipitation-Western analysis revealed a dramatically enhanced complex formation between CK1delta and tubulins from mitotic extracts after introducing DNA damage. The high affinity of CK1delta to the Spindle Apparatus in DNA-damaged cells and its ability to phosphorylate several microtubule-associated proteins points to a regulatory role of CK1delta at mitosis.
Lars Behrend - One of the best experts on this subject based on the ideXlab platform.
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interaction of casein kinase 1 delta ck1delta with post golgi structures microtubules and the Spindle Apparatus
European Journal of Cell Biology, 2000Co-Authors: Lars Behrend, Jochen Heukeshoven, Gabriel Rutter, Wolfgang Deppert, Marion Kurth, Martin Stoter, Uwe KnippschildAbstract:Summary Members of the casein kinase 1 family of serine/threonine kinases are highly conserved from yeast to mammals and seem to play an important role in vesicular trafficking, DNA repair, cell cycle progression and cytokinesis. We here report that in interphase cells of various mammalian species casein kinase 1 delta (CK1δ) specifically interacts with the trans Golgi network and cytoplasmic, granular particles that associate with microtubules. Furthermore, at mitosis CK1δ is recruited to the Spindle Apparatus and the centrosomes in cells, which have been exposed to DNA-damaging agents like etoposide or γ-irradiation. In addition, determination of the affinity of CK1δ to different tubulin isoforms in immunoprecipitation-Western analysis revealed a dramatically enhanced complex formation between CK1δ and tubulins from mitotic extracts after introducing DNA damage. The high affinity of CK1δ to the Spindle Apparatus in DNA-damaged cells and its ability to phosphorylate several microtubule-associated proteins points to a regulatory role of CK1δ at mitosis.
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interaction of casein kinase 1 delta ck1delta with post golgi structures microtubules and the Spindle Apparatus
European Journal of Cell Biology, 2000Co-Authors: Lars Behrend, Jochen Heukeshoven, Gabriel Rutter, Wolfgang Deppert, Marion Kurth, Martin Stoter, Uwe KnippschildAbstract:Members of the casein kinase 1 family of serine/threonine kinases are highly conserved from yeast to mammals and seem to play an important role in vesicular trafficking, DNA repair, cell cycle progression and cytokinesis. We here report that in interphase cells of various mammalian species casein kinase 1 delta (CK1delta) specifically interacts with the trans Golgi network and cytoplasmic, granular particles that associate with microtubules. Furthermore, at mitosis CK1delta is recruited to the Spindle Apparatus and the centrosomes in cells, which have been exposed to DNA-damaging agents like etoposide or gammairradiation. In addition, determination of the affinity of CK1delta to different tubulin isoforms in immunoprecipitation-Western analysis revealed a dramatically enhanced complex formation between CK1delta and tubulins from mitotic extracts after introducing DNA damage. The high affinity of CK1delta to the Spindle Apparatus in DNA-damaged cells and its ability to phosphorylate several microtubule-associated proteins points to a regulatory role of CK1delta at mitosis.
Erich A Nigg - One of the best experts on this subject based on the ideXlab platform.
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novel asymmetrically localizing components of human centrosomes identified by complementary proteomics methods
The EMBO Journal, 2011Co-Authors: Lis Jakobsen, K Vanselow, Marie Skogs, Yusuke Toyoda, Emma Lundberg, Ina Poser, Lasse Gaarde Falkenby, Martin V Bennetzen, Jens Westendorf, Erich A NiggAbstract:Centrosomes in animal cells are dynamic organelles with a proteinaceous matrix of pericentriolar material assembled around a pair of centrioles. They organize the microtubule cytoskeleton and the mitotic Spindle Apparatus. Mature centrioles are essential for biogenesis of primary cilia that mediate key signalling events. Despite recent advances, the molecular basis for the plethora of processes coordinated by centrosomes is not fully understood. We have combined protein identification and localization, using PCP-SILAC mass spectrometry, BAC transgeneOmics, and antibodies to define the constituents of human centrosomes. From a background of non-specific proteins, we distinguished 126 known and 40 candidate centrosomal proteins, of which 22 were confirmed as novel components. An antibody screen covering 4000 genes revealed an additional 113 candidates. We illustrate the power of our methods by identifying a novel set of five proteins preferentially associated with mother or daughter centrioles, comprising genes implicated in cell polarity. Pulsed labelling demonstrates a remarkable variation in the stability of centrosomal protein complexes. These spatiotemporal proteomics data provide leads to the further functional characterization of centrosomal proteins.
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human tpx2 is required for targeting aurora a kinase to the Spindle
Journal of Cell Biology, 2002Co-Authors: Thomas A Kufer, Herman H W Sillje, Roman Korner, Oliver J Gruss, Patrick Meraldi, Erich A NiggAbstract:Aurora-A is a serine-threonine kinase implicated in the assembly and maintenance of the mitotic Spindle. Here we show that human Aurora-A binds to TPX2, a prominent component of the Spindle Apparatus. TPX2 was identified by mass spectrometry as a major protein coimmunoprecipitating specifically with Aurora-A from mitotic HeLa cell extracts. Conversely, Aurora-A could be detected in TPX2 immunoprecipitates. This indicates that subpopulations of these two proteins undergo complex formation in vivo. Binding studies demonstrated that the NH2 terminus of TPX2 can directly interact with the COOH-terminal catalytic domain of Aurora-A. Although kinase activity was not required for this interaction, TPX2 was readily phosphorylated by Aurora-A. Upon siRNA-mediated elimination of TPX2 from cells, the association of Aurora-A with the Spindle microtubules was abolished, although its association with Spindle poles was unaffected. Conversely, depletion of Aurora-A by siRNA had no detectable influence on the localization of TPX2. We propose that human TPX2 is required for targeting Aurora-A kinase to the Spindle Apparatus. In turn, Aurora-A might regulate the function of TPX2 during Spindle assembly.
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dynamic changes in nuclear architecture during mitosis on the role of protein phosphorylation in Spindle assembly and chromosome segregation
Experimental Cell Research, 1996Co-Authors: Erich A Nigg, Anne Blangy, Heidi A LaneAbstract:During mitosis, the vertebrate cell nucleus undergoes profound changes in architecture. At the onset of mitosis, the nuclear envelope breaks down, the nuclear lamina is depolymerized, and interphase chromatin is condensed to chromosomes. Concomitantly, cytoplasmic microtubules are reorganized into a mitotic Spindle Apparatus, a highly dynamic structure required for the segregation of sister chromatids. Many of the above events are controlled by reversible phosphorylation. Hence, our laboratory is interested in characterizing the kinases involved in promoting progression through mitosis and in identifying their relevant substrates. Prominent among the kinases responsible for regulating entry into mitosis is the Cdc2 kinase, the first member of the cyclin dependent kinase (Cdk) family. Recently, we found that Cdc2 phosphorylates HsEg5, a human kinesin-related motor protein associated with centrosomes and the Spindle Apparatus. Our results indicate that phosphorylation regulates the association of HsEg5 with the mitotic Spindle and that the function of this plus-end directed motor is essential for centrosome separation and bipolar Spindle formation. Another kinase implicated in regulating progression through mitosis is Plk1 (polo-like kinase 1), the human homologue of the Drosophila gene product "polo." By antibody microinjection we have found that Plk1 is required for the functional maturation of centrosomes and hence for entry into mitosis. Furthermore, we found that microinjected anti-Plk1 antibodies caused a more severe block to cell cycle progression in diploid fibroblasts than in immortalized tumor cells. This observation hints at the existence of a checkpoint linking Cdc2 activation to the presence of functional centrosomes.
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phosphorylation by p34cdc2 regulates Spindle association of human eg5 a kinesin related motor essential for bipolar Spindle formation in vivo
Cell, 1995Co-Authors: Anne Blangy, Heidi Lane, Pierre Dherin, Maryannick Harper, Michel Kress, Erich A NiggAbstract: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 p34cd2/ 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 p34cd2 protein kinase directly regulates its localization.
Gabriel Rutter - One of the best experts on this subject based on the ideXlab platform.
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interaction of casein kinase 1 delta ck1delta with post golgi structures microtubules and the Spindle Apparatus
European Journal of Cell Biology, 2000Co-Authors: Lars Behrend, Jochen Heukeshoven, Gabriel Rutter, Wolfgang Deppert, Marion Kurth, Martin Stoter, Uwe KnippschildAbstract:Summary Members of the casein kinase 1 family of serine/threonine kinases are highly conserved from yeast to mammals and seem to play an important role in vesicular trafficking, DNA repair, cell cycle progression and cytokinesis. We here report that in interphase cells of various mammalian species casein kinase 1 delta (CK1δ) specifically interacts with the trans Golgi network and cytoplasmic, granular particles that associate with microtubules. Furthermore, at mitosis CK1δ is recruited to the Spindle Apparatus and the centrosomes in cells, which have been exposed to DNA-damaging agents like etoposide or γ-irradiation. In addition, determination of the affinity of CK1δ to different tubulin isoforms in immunoprecipitation-Western analysis revealed a dramatically enhanced complex formation between CK1δ and tubulins from mitotic extracts after introducing DNA damage. The high affinity of CK1δ to the Spindle Apparatus in DNA-damaged cells and its ability to phosphorylate several microtubule-associated proteins points to a regulatory role of CK1δ at mitosis.
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interaction of casein kinase 1 delta ck1delta with post golgi structures microtubules and the Spindle Apparatus
European Journal of Cell Biology, 2000Co-Authors: Lars Behrend, Jochen Heukeshoven, Gabriel Rutter, Wolfgang Deppert, Marion Kurth, Martin Stoter, Uwe KnippschildAbstract:Members of the casein kinase 1 family of serine/threonine kinases are highly conserved from yeast to mammals and seem to play an important role in vesicular trafficking, DNA repair, cell cycle progression and cytokinesis. We here report that in interphase cells of various mammalian species casein kinase 1 delta (CK1delta) specifically interacts with the trans Golgi network and cytoplasmic, granular particles that associate with microtubules. Furthermore, at mitosis CK1delta is recruited to the Spindle Apparatus and the centrosomes in cells, which have been exposed to DNA-damaging agents like etoposide or gammairradiation. In addition, determination of the affinity of CK1delta to different tubulin isoforms in immunoprecipitation-Western analysis revealed a dramatically enhanced complex formation between CK1delta and tubulins from mitotic extracts after introducing DNA damage. The high affinity of CK1delta to the Spindle Apparatus in DNA-damaged cells and its ability to phosphorylate several microtubule-associated proteins points to a regulatory role of CK1delta at mitosis.
Jochen Heukeshoven - One of the best experts on this subject based on the ideXlab platform.
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interaction of casein kinase 1 delta ck1delta with post golgi structures microtubules and the Spindle Apparatus
European Journal of Cell Biology, 2000Co-Authors: Lars Behrend, Jochen Heukeshoven, Gabriel Rutter, Wolfgang Deppert, Marion Kurth, Martin Stoter, Uwe KnippschildAbstract:Summary Members of the casein kinase 1 family of serine/threonine kinases are highly conserved from yeast to mammals and seem to play an important role in vesicular trafficking, DNA repair, cell cycle progression and cytokinesis. We here report that in interphase cells of various mammalian species casein kinase 1 delta (CK1δ) specifically interacts with the trans Golgi network and cytoplasmic, granular particles that associate with microtubules. Furthermore, at mitosis CK1δ is recruited to the Spindle Apparatus and the centrosomes in cells, which have been exposed to DNA-damaging agents like etoposide or γ-irradiation. In addition, determination of the affinity of CK1δ to different tubulin isoforms in immunoprecipitation-Western analysis revealed a dramatically enhanced complex formation between CK1δ and tubulins from mitotic extracts after introducing DNA damage. The high affinity of CK1δ to the Spindle Apparatus in DNA-damaged cells and its ability to phosphorylate several microtubule-associated proteins points to a regulatory role of CK1δ at mitosis.
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interaction of casein kinase 1 delta ck1delta with post golgi structures microtubules and the Spindle Apparatus
European Journal of Cell Biology, 2000Co-Authors: Lars Behrend, Jochen Heukeshoven, Gabriel Rutter, Wolfgang Deppert, Marion Kurth, Martin Stoter, Uwe KnippschildAbstract:Members of the casein kinase 1 family of serine/threonine kinases are highly conserved from yeast to mammals and seem to play an important role in vesicular trafficking, DNA repair, cell cycle progression and cytokinesis. We here report that in interphase cells of various mammalian species casein kinase 1 delta (CK1delta) specifically interacts with the trans Golgi network and cytoplasmic, granular particles that associate with microtubules. Furthermore, at mitosis CK1delta is recruited to the Spindle Apparatus and the centrosomes in cells, which have been exposed to DNA-damaging agents like etoposide or gammairradiation. In addition, determination of the affinity of CK1delta to different tubulin isoforms in immunoprecipitation-Western analysis revealed a dramatically enhanced complex formation between CK1delta and tubulins from mitotic extracts after introducing DNA damage. The high affinity of CK1delta to the Spindle Apparatus in DNA-damaged cells and its ability to phosphorylate several microtubule-associated proteins points to a regulatory role of CK1delta at mitosis.