Centrosome organization and centriole architecture: Their sensitivity to divalent cations

Paintrand, M; Moudjou, Mohammed; Delacroix, Hervé; Bornens, Michel

doi:10.1016/1047-8477(92)90011-x

Cited by 345 publications

(316 citation statements)

References 37 publications

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“…For this purpose we were using the technique of whole mount preparation of isolated centrosomes and in vitro nucleation of microtubules. Such an analysis of isolated centrosomes (as opposed to in situ) can be extremely informative in revealing structural features of this organelle in more detail (see the work of Chretien et al 1997;Lange and Gull 1996b;Moritz et al 1995a;Paintrand et al 1992).…”

Section: Discussionmentioning

confidence: 99%

“…Compared to centrosomes isolated from mammalian cells, Drosophila early embryonic centrosomes have an average size of about 0.75 lm as compared to about 0.5 lm for the typical mammalian centrosome. This size difference is due to the low amount of pericentriolar material in somatic mammalian cells and is illustrated by the fact that centrioles can easily be visualized in the mammalian centrosome (Komesli et al 1989;Lange and Gull 1996b;Paintrand et al 1992) also by whole mount techniques while the centriole from the early Drosophila embryo (as shown in our data) is almost impossible to discern concealed by the abundant pericentriolar material (compare also to TEM images of sections of isolated Drosophila centrosomes in Lange et al (2000) and Moritz et al (1995a). Debec et al (1999) characterized Drosophila early embryo centrosomes in situ through energy filtering TEM and observed in interphase centrosomes, structurally compact pericentriolar material that displayed long protrusion nucleating relatively few microtubules.…”

Section: Discussionmentioning

confidence: 99%

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Structure and microtubule-nucleation activity of isolated Drosophila embryo centrosomes characterized by whole mount scanning and transmission electron microscopy

Lange

Kirfel

Gestmann

et al. 2005

Histochem Cell Biol

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Experimental approaches in Drosophila melanogaster over the last 20 years have played a fundamental role in elucidating the function, structure and molecular composition of the centrosome. However, quantitative data on the structure and function of the Drosophila centrosome are still lacking. This study uses, for the first time, whole mount electron microscopy in combination with negative staining on isolated centrosomes from the early Drosophila embryos to analyze its dimensions, structure and capacity to nucleate microtubules in vitro. We show that these organelles are on average 0.75 lm in diameter and have abundant pericentriolar material which often appears fibrillar and with bulbous protrusions. Corresponding to the abundant pericentriolar material, extensive microtubule nucleation occurs. Quantification of the number of microtubules nucleated showed that 50-300 active nucleation sites are present. We examined via electron microscopy immunogold labeling the distribution of c-tubulin, CNN, Asp and the MPM-2 epitopes that are phosphorylated through Polo and the Cdk1 kinase. The distribution of these proteins is homogeneous, with the MPM-2 epitopes exhibiting the highest density. In contrast, centrosomal subdomains are identified using a centriole marker to relate centrosome size to the centriole number by electron microscopy. In conclusion, we present a clear-cut technique assaying and quantifying the microtubule nucleation capacity and antigen distribution complementing molecular studies on centrosome protein complexes, cell organelle assembly and protein composition.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Structure and microtubule-nucleation activity of isolated Drosophila embryo centrosomes characterized by whole mount scanning and transmission electron microscopy

Lange

Kirfel

Gestmann

et al. 2005

Histochem Cell Biol

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“…CPAP also cooperates with additional proteins, including CP110, in determining the length of nascent centrioles (Kohlmaier et al , 2009; Schmidt et al , 2009; Tang et al , 2009; Comartin et al , 2013; Lin et al , 2013; Sharma et al , 2016). Finally, centriole maturation is completed by the acquisition of subdistal and distal appendages (Paintrand et al , 1992; Tateishi et al , 2013). …”

Section: Introductionmentioning

confidence: 99%

Quantitative analysis of human centrosome architecture by targeted proteomics and fluorescence imaging

et al. 2016

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Centrioles are essential for the formation of centrosomes and cilia. While numerical and/or structural centrosomes aberrations are implicated in cancer, mutations in centriolar and centrosomal proteins are genetically linked to ciliopathies, microcephaly, and dwarfism. The evolutionarily conserved mechanisms underlying centrosome biogenesis are centered on a set of key proteins, including Plk4, Sas‐6, and STIL, whose exact levels are critical to ensure accurate reproduction of centrioles during cell cycle progression. However, neither the intracellular levels of centrosomal proteins nor their stoichiometry within centrosomes is presently known. Here, we have used two complementary approaches, targeted proteomics and EGFP‐tagging of centrosomal proteins at endogenous loci, to measure protein abundance in cultured human cells and purified centrosomes. Our results provide a first assessment of the absolute and relative amounts of major components of the human centrosome. Specifically, they predict that human centriolar cartwheels comprise up to 16 stacked hubs and 1 molecule of STIL for every dimer of Sas‐6. This type of quantitative information will help guide future studies of the molecular basis of centrosome assembly and function.

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“…Malfunction in these processes may lead to the development of tumors (Brinkley and Goepfert, 1998;Doxsey, 1998;Salisbury et al, 1999). A previous study indicates that a proteinaceous linker between the pair of centrioles mediates their cohesion (Paintrand et al, 1992), though the identity of this linker remains unknown. Recent studies demonstrate that several proteins, including NIMA-related kinase 2 (Nek2), protein phosphatase 1 (PP1), C-Nap1, dynamin 2, and RanBP1, are involved in centriolar cohesion (Mayor et al, 2000;Meraldi and Nigg, 2001;Di Fiore et al, 2003;Thompson et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

Rootletin Interacts with C-Nap1 and May Function as a Physical Linker between the Pair of Centrioles/Basal Bodies in Cells

Yang

Adamian

2006

MBoC

143

174

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Rootletin, a major structural component of the ciliary rootlet, is located at the basal bodies and centrosomes in ciliated and nonciliated cells, respectively. Here we investigated its potential role in the linkage of basal bodies/centrioles and the mechanism involved in such linkages. We show that rootletin interacts with C-Nap1, a protein restricted at the ends of centrioles and functioning in centrosome cohesion in interphase cells. Their interaction in vivo is supported by their colocalization at the basal bodies/centrioles and coordinated association with the centrioles during the cell cycle. Ultrastructural examinations demonstrate that rootletin fibers connect the basal bodies in ciliated cells and are present both at the ends of and in between the pair of centrioles in nonciliated cells. The latter finding stands in contrast with C-Nap1, which is present only at the ends of the centrioles. Transient expression of C-Nap1 fragments dissociated rootletin fibers from the centrioles, resulting in centrosome separation in interphase. Overexpression of rootletin in cells caused multinucleation, micronucleation, and irregularity of nuclear shape and size, indicative of defects in chromosome separation. These data suggest that rootletin may function as a physical linker between the pair of basal bodies/centrioles by binding to C-Nap1.

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Centrosome organization and centriole architecture: Their sensitivity to divalent cations

Cited by 345 publications

References 37 publications

Structure and microtubule-nucleation activity of isolated Drosophila embryo centrosomes characterized by whole mount scanning and transmission electron microscopy

Structure and microtubule-nucleation activity of isolated Drosophila embryo centrosomes characterized by whole mount scanning and transmission electron microscopy

Quantitative analysis of human centrosome architecture by targeted proteomics and fluorescence imaging

Rootletin Interacts with C-Nap1 and May Function as a Physical Linker between the Pair of Centrioles/Basal Bodies in Cells

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