Filamentous polymers induced by overexpression of a novel centrosomal protein, Cep135

Ryu, Jung Hwa; Essner, Russell; Ohta, Toshiro; Kuriyama, Ryoko

doi:10.1002/(sici)1097-0029(20000601)49:5<478::aid-jemt10>3.0.co;2-j

Cited by 8 publications

(6 citation statements)

References 29 publications

(30 reference statements)

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“…As we know, overexpression of Cep135, a novel structural component of the centrosome, can lead to filamentous aggregates in and outside the centrosome. 39 Similar to the result of Cep135 overexpression, both centrosomal protein RanBPM 40 and g-tubulin 41 can cause ectopic microtubule nucleation, namely, can cause formation of extra MTOCs. The ability of Ceap to induce extra MTOCs formation would be through the mechanism that Ceap could promote the mutation of g-tubulin and subsequently would lead to aggregation of g-tubulin.…”

Section: Discussionmentioning

confidence: 76%

Characterization of Ceap-11 and Ceap-16, Two Novel Splicing–Variant–Proteins, Associated with Centrosome, Microtubule Aggregation and Cell Proliferation

Wang

Huang

et al. 2004

Journal of Molecular Biology

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Section: Discussionmentioning

confidence: 76%

Characterization of Ceap-11 and Ceap-16, Two Novel Splicing–Variant–Proteins, Associated with Centrosome, Microtubule Aggregation and Cell Proliferation

Wang

Huang

et al. 2004

Journal of Molecular Biology

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“…Samples for electron microscopy were prepared according to previously described procedure (Ryu et al, 2000). Briefly, CHO cells, cultured in a plastic culture dish were transfected with the GFPtagged CHO1FЈ and synchronized at the stage of cytokinesis.…”

Section: Electron Microscopymentioning

confidence: 99%

Kinesin-like Protein CHO1 Is Required for the Formation of Midbody Matrix and the Completion of Cytokinesis in Mammalian Cells

Matulienė

Kuriyama

2002

MBoC

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137

121

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CHO1 is a mammalian kinesin-like motor protein of the MKLP1 subfamily. It associates with the spindle midzone during anaphase and concentrates to a midbody matrix during cytokinesis. CHO1 was originally implicated in karyokinesis, but the invertebrate homologues of CHO1 were shown to function in the midzone formation and cytokinesis. To analyze the role of the protein in mammalian cells, we mutated the ATP-binding site of CHO1 and expressed it in CHO cells. Mutant protein (CHO1F') was able to interact with microtubules via ATP-independent microtubule-binding site(s) but failed to accumulate at the midline of the central spindle and affected the localization of endogenous CHO1. Although the segregation of chromosomes, the bundling of midzone microtubules, and the initiation of cytokinesis proceeded normally in CHO1F'-expressing cells, the completion of cytokinesis was inhibited. Daughter cells were frequently entering interphase while connected by a microtubule-containing cytoplasmic bridge from which the dense midbody matrix was missing. Depletion of endogenous CHO1 via RNA-mediated interference also affected the formation of midbody matrix in dividing cells, caused the disorganization of midzone microtubules, and resulted in abortive cytokinesis. Thus, CHO1 may not be required for karyokinesis, but it is essential for the proper midzone/midbody formation and cytokinesis in mammalian cells.

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“…Cells expressing GFP-tagged Cep135 polypeptides were marked with a diamond scribe and further processed for thin section electron microscopy (Ryu et al, 2000). For immunoelectron microscopy, we used a protocol for preembedding immunogold staining (Kuriyama, 1989;Ryu et al, 2000). In brief, CHO cells in a culture chamber were fixed with 3% formaldehyde in PEM (100 mM Pipes at pH 6.8, 1 mM EGTA, 1 mM MgCl 2 ) for 10 min at room temperature.…”

Section: Electron Microscopymentioning

confidence: 99%

Section: Electron Microscopymentioning

confidence: 99%

Characterization of Cep135, a novel coiled-coil centrosomal protein involved in microtubule organization in mammalian cells

Ohta

Essner

Ryu

et al. 2002

The Journal of Cell Biology

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124

122

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By using monoclonal antibodies raised against isolated clam centrosomes, we have identified a novel 135-kD centrosomal protein (Cep135), present in a wide range of organisms. Cep135 is located at the centrosome throughout the cell cycle, and localization is independent of the microtubule network. It distributes throughout the centrosomal area in association with the electron-dense material surrounding centrioles. Sequence analysis of cDNA isolated from CHO cells predicted a protein of 1,145–amino acid residues with extensive α-helical domains. Expression of a series of deletion constructs revealed the presence of three independent centrosome-targeting domains. Overexpression of Cep135 resulted in the accumulation of unique whorl-like particles in both the centrosome and the cytoplasm. Although their size, shape, and number varied according to the level of protein expression, these whorls were composed of parallel dense lines arranged in a 6-nm space. Altered levels of Cep135 by protein overexpression and/or suppression of endogenous Cep135 by RNA interference caused disorganization of interphase and mitotic spindle microtubules. Thus, Cep135 may play an important role in the centrosomal function of organizing microtubules in mammalian cells.

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Filamentous polymers induced by overexpression of a novel centrosomal protein, Cep135

Cited by 8 publications

References 29 publications

Characterization of Ceap-11 and Ceap-16, Two Novel Splicing–Variant–Proteins, Associated with Centrosome, Microtubule Aggregation and Cell Proliferation

Characterization of Ceap-11 and Ceap-16, Two Novel Splicing–Variant–Proteins, Associated with Centrosome, Microtubule Aggregation and Cell Proliferation

Kinesin-like Protein CHO1 Is Required for the Formation of Midbody Matrix and the Completion of Cytokinesis in Mammalian Cells

Characterization of Cep135, a novel coiled-coil centrosomal protein involved in microtubule organization in mammalian cells

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