Cell cycle-dependent, in vitro assembly of microtubules onto pericentriolar material of HeLa cells.

Telzer, Bruce R.; Rosenbaum, Joel L.

doi:10.1083/jcb.81.3.484

Cited by 97 publications

(53 citation statements)

References 15 publications

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“…Various investigators have shown that PCM is still present and functionally active in mammalian cells after cells were lysed in buffers containing 0.25-0.5% Triton X-100 (15,18,30). To obtain images of centrosomes and polar regions that are more clearly outlined than in intact cells, spermatocyte preparations were subjected to different lysis media in another set of experiments.…”

Section: Resultsmentioning

confidence: 99%

“…Electron microscopical investigations show that normal centrosomes are compound structures composed of a centriole pair, a cloud of darkly staining pericentriolar material (PCM),' and microtubules radiating from it ("aster" microtubules). It became evident that the capacity of the centrosome to nucleate microtubules is due to the PCM (l 5,30), and the PCM is regarded as the important component for spindle pole formation, while the centrioles do not seem to play a critical role during this process (3,5,6,17). The presence of PCM could recently even be verified at the spindle poles in plant cells, a well known example of acentric spindle formation (8,32).…”

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confidence: 99%

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Aster-free spindle poles in insect spermatocytes: evidence for chromosome-induced spindle formation?

Walter¹,

Fuge²,

Dietz³

et al. 1986

The Journal of Cell Biology

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Abstract. Tipulid spermatocytes form normally functioning bipolar spindles after one of the centrosomes is experimentally dislocated from the nucleus in late diakinesis (Dietz, R., 1959, Z. Naturforsch., 14b:749-752; Dietz, R., 1963, Zool. Anz. Suppl., 23:131-138; Dietz, R., 1966, Heredity, 19:161-166). The possibility that dissociated pericentriolar material (PCM) is nevertheless responsible for the formation of the spindle in these cells cannot be ruled out based on live observation. In studying serial sections of complete cells and of lysed cells, it was found that centrosomefree spindle poles in the crane fly show neither pericentriolar-like material nor aster microtubules, whereas the displaced centrosomes appear complete, i.e., consist of a centriole pair, aster microtubules, and PCM. Exposure to a lysis buffer containing tubulin resulted in an increase of centrosomal asters due to aster micotubule polymerization. Aster-free spindle poles did not show any reaction, also indicating the absence of PCM at these poles. The results favor the hypothesis of chromosome-induced spindle pole formation at the onset of prometaphase and the dispensability of PCM in Pales.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Aster-free spindle poles in insect spermatocytes: evidence for chromosome-induced spindle formation?

Walter¹,

Fuge²,

Dietz³

et al. 1986

The Journal of Cell Biology

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“…Isolated centrosomes radially nucleate microtubules forming asterlike structures [McGill and Brinkley, 1975;Snyder and McIntosh, 1975;Weisenberg and Rosenfeld, 1975;Gould and Borisy, 1977;Pepper and Brinkley, 1979;Telzer and Rosenbaum, 1979;Bergen et al, 19801. On the other hand, the basal body, having the same structure as the centriole, nucleates microtubules in vitro at either end of it in a template manner [Snell et al, 19741.…”

Section: Introductionmentioning

confidence: 99%

51‐kd protein, a component of microtubule‐organizing granules in the mitotic apparatus involved in aster formation in vitro

Toriyama¹,

Ohta²,

Endo³

et al. 1988

Cell Motil. Cytoskeleton

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Mitotic apparatuses (MAS) isolated from sea urchin metaphase eggs were chilled on ice to depolymerize microtubules, homogenized, and incubated with tubulin. This caused formation of many small asters with microtubules focusing on granules which were probably fragments of the centrosome. The aster-forming protein components of the granules in the homogenized MAS were solubilized in 0.5 M KCI containing SO % glycerol. After dialysis against low-ionic-strength buffer solution, proteins congregated to form granular assembly capable of initiating aster formation. Phosphocellulose column chromatography enabled the separation of the aster-forming protein fraction which contained a S1,OOO molecular weight protein (51-kd protein) as a major component. The protein fraction possessing the aster-forming activity was also prepared from methaphase whole egg homogenate, and the elution profile of the 51-kd protein on phosphocellulose column also coincided with that of the aster-forming activity. The granular assembly reconstituted from the phosphocellulose fraction formed asters whose microtubules show the same growth rate and length distribution as those of asters reconstructed from the granules in the homogenized MAS. Anti-51-kd protein antibody that was raised in rabbit and affinity-purified stained the center of asters which were reconstructed either from the granules in the homogenized MAS or from the granular assembly reconstituted from the phosphocellulose fraction. These results suggest that the S1-kd protein is a component in the aster-forming activity of the centrosomal component in vitro.

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“…It is generally believed that these organelles, or closely associated structures (the satellite material surrounding centrioles ; the basal plate of cilia), determine the organization of microtubules into cytoplasmic networks, mitotic spindles, cilia, and flagella. Attempts to isolate centrioles and basal bodies, however, have met with only limited success (e.g ., 33,37) . Thus the replication of centrioles and the maturation of basal bodies has been elegantly described at the ultrastructural level (2, 20) but has not been studied biochemically; an incompletely characterized ATPase appears to be the only enzyme that has been localized to basal bodies (1); and the protein composition of pericentriolar material is unknown except for the recent demonstration in a highly specialized cnidarian sperm of pericentriolar actin (21).…”

Section: Discussionmentioning

confidence: 99%

Purine nucleoside phosphorylase is associated with centrioles and basal bodies.

Oliver¹,

Osborne²,

Pfeiffer³

et al. 1981

The Journal of Cell Biology

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We have localized a fraction of the enzyme, purine nucleoside phosphorylase (PNP), to the centrioles and basal bodies of mammalian, avian, and protozoan cells. Two completely independent methods were used, one based on the ultrastructural cytochemistry of the enzyme activity and one based on immunofluorescence microscopy using an antibody raised in rabbit against purified human PNP. PNP catalyzes the reversible conversion of purine nucleosides and inorganic phosphate to the corresponding purine bases and ribose-1-phosphate . Its partial localization to centrioles and basal bodies raises the possibility that purine compounds are involved in centriole replication and/or in the regulation of microtubule assembly in vivo .No centriolar PNP could be detected in primary skin fibroblast from two infants with severe immunodeficiency disease associated with the absence of soluble PNP . This raises the possibility that defects in centriole function may contribute to the impaired division and maturation of T lymphoid precursor in this inherited disorder .Initially, the immunofluorescence analyses were complicated by a residual centriole-binding antibody that persisted in immunoglobulins from immune animals after complete removal of anti-PNP by affinity chromatography . Binding was abolished by exposure of cells to sodium periodate, indicating that this (and possibly other) "spontaneous" anticentriole antibodies in rabbit serum may be directed against carbohydrates .It has become clear from ultrastructural studies that the majority of microtubules in animal cells grow from distinct regions known as microtubule-organizing centers (MTOCs). These structures, which consist of centrioles and their associated satellite substance, basal bodies, and kinetochores, may be the key to understanding the organization and assembly of cytoplasmic, mitotic spindle, and ciliary microtubules . Nevertheless, little is known of their biochemical composition or function (10, 38).We report here that an enzyme, purine nucleoside phosphorylase (PNP ; purine nucleoside ; orthophosphate ribosyltransferase, EC 2 .4 .2.1) is associated with centrioles and their derivatives, the basal bodies of ciliated cells . This enzyme, which catalyzes the reversible reaction of purine nucleosides (guanosine, inosine, and xanthosine, and their deoxyhomologues) with inorganic phosphate to form free bases and (deoxy)ribose-1-phosphate, has been measured previously in soluble extracts of cells ranging from bacteria to man . Its forward reaction (nu-

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Cell cycle-dependent, in vitro assembly of microtubules onto pericentriolar material of HeLa cells.

Cited by 97 publications

References 15 publications

Aster-free spindle poles in insect spermatocytes: evidence for chromosome-induced spindle formation?

Aster-free spindle poles in insect spermatocytes: evidence for chromosome-induced spindle formation?

51‐kd protein, a component of microtubule‐organizing granules in the mitotic apparatus involved in aster formation in vitro

Purine nucleoside phosphorylase is associated with centrioles and basal bodies.

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