We identified Xenopus pericentriolar material-1 (PCM-1), which had been reported to constitute pericentriolar material, cloned its cDNA, and generated a specific pAb against this molecule. Immunolabeling revealed that PCM-1 was not a pericentriolar material protein, but a specific component of centriolar satellites, morphologically characterized as electron-dense granules, ∼70–100 nm in diameter, scattered around centrosomes. Using a GFP fusion protein with PCM-1, we found that PCM-1–containing centriolar satellites moved along microtubules toward their minus ends, i.e., toward centrosomes, in live cells, as well as in vitro reconstituted asters. These findings defined centriolar satellites at the molecular level, and explained their pericentriolar localization. Next, to understand the relationship between centriolar satellites and centriolar replication, we examined the expression and subcellular localization of PCM-1 in ciliated epithelial cells during ciliogenesis. When ciliogenesis was induced in mouse nasal respiratory epithelial cells, PCM-1 immunofluorescence was markedly elevated at the apical cytoplasm. At the electron microscopic level, anti–PCM-1 pAb exclusively labeled fibrous granules, but not deuterosomes, both of which have been suggested to play central roles in centriolar replication in ciliogenesis. These findings suggested that centriolar satellites and fibrous granules are identical novel nonmembranous organelles containing PCM-1, which may play some important role(s) in centriolar replication.
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29 November 1999
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November 29 1999
Centriolar Satellites: Molecular Characterization, Atp-Dependent Movement toward Centrioles and Possible Involvement in Ciliogenesis
Akiharu Kubo,
Akiharu Kubo
aTsukita Cell Axis Project, Exploratory Research for Advanced Technology, Japan Science and Technology Corporation, Kyoto Research Park, Shimogyo-ku, Kyoto 600-8813, Japan
bDepartment of Cell Biology, Kyoto University Faculty of Medicine, Sakyo-ku, Kyoto 606-8501, Japan
cDepartment of Dermatology, Osaka University School of Medicine, Suita 565-0871, Japan
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Hiroyuki Sasaki,
Hiroyuki Sasaki
dLaboratory of Cell Biology, KAN Research Institute Inc., Kyoto Research Park, Shimogyo-ku, Kyoto 600-8815, Japan
eDepartment of Molecular Cell Biology, Institute of DNA Medicine, Jikei University School of Medicine, Minato-ku, Tokyo 105-0003, Japan
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Akiko Yuba-Kubo,
Akiko Yuba-Kubo
aTsukita Cell Axis Project, Exploratory Research for Advanced Technology, Japan Science and Technology Corporation, Kyoto Research Park, Shimogyo-ku, Kyoto 600-8813, Japan
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Shoichiro Tsukita,
Shoichiro Tsukita
aTsukita Cell Axis Project, Exploratory Research for Advanced Technology, Japan Science and Technology Corporation, Kyoto Research Park, Shimogyo-ku, Kyoto 600-8813, Japan
bDepartment of Cell Biology, Kyoto University Faculty of Medicine, Sakyo-ku, Kyoto 606-8501, Japan
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Nobuyuki Shiina
Nobuyuki Shiina
aTsukita Cell Axis Project, Exploratory Research for Advanced Technology, Japan Science and Technology Corporation, Kyoto Research Park, Shimogyo-ku, Kyoto 600-8813, Japan
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Akiharu Kubo
aTsukita Cell Axis Project, Exploratory Research for Advanced Technology, Japan Science and Technology Corporation, Kyoto Research Park, Shimogyo-ku, Kyoto 600-8813, Japan
bDepartment of Cell Biology, Kyoto University Faculty of Medicine, Sakyo-ku, Kyoto 606-8501, Japan
cDepartment of Dermatology, Osaka University School of Medicine, Suita 565-0871, Japan
Hiroyuki Sasaki
dLaboratory of Cell Biology, KAN Research Institute Inc., Kyoto Research Park, Shimogyo-ku, Kyoto 600-8815, Japan
eDepartment of Molecular Cell Biology, Institute of DNA Medicine, Jikei University School of Medicine, Minato-ku, Tokyo 105-0003, Japan
Akiko Yuba-Kubo
aTsukita Cell Axis Project, Exploratory Research for Advanced Technology, Japan Science and Technology Corporation, Kyoto Research Park, Shimogyo-ku, Kyoto 600-8813, Japan
Shoichiro Tsukita
aTsukita Cell Axis Project, Exploratory Research for Advanced Technology, Japan Science and Technology Corporation, Kyoto Research Park, Shimogyo-ku, Kyoto 600-8813, Japan
bDepartment of Cell Biology, Kyoto University Faculty of Medicine, Sakyo-ku, Kyoto 606-8501, Japan
Nobuyuki Shiina
aTsukita Cell Axis Project, Exploratory Research for Advanced Technology, Japan Science and Technology Corporation, Kyoto Research Park, Shimogyo-ku, Kyoto 600-8813, Japan
The online version of this article contains supplemental material.
Abbreviations used in this paper: aa, amino acids; AMP-PNP, adenylylimido diphosphate; GFP, green fluorescent protein; MT, microtubule; ORF, open reading frame; pAb, polyclonal antibody; PCM-1, pericentriolar material-1.
Received:
July 29 1999
Revision Requested:
September 28 1999
Accepted:
October 14 1999
Online ISSN: 1540-8140
Print ISSN: 0021-9525
© 1999 The Rockefeller University Press
1999
The Rockefeller University Press
J Cell Biol (1999) 147 (5): 969–980.
Article history
Received:
July 29 1999
Revision Requested:
September 28 1999
Accepted:
October 14 1999
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Citation
Akiharu Kubo, Hiroyuki Sasaki, Akiko Yuba-Kubo, Shoichiro Tsukita, Nobuyuki Shiina; Centriolar Satellites: Molecular Characterization, Atp-Dependent Movement toward Centrioles and Possible Involvement in Ciliogenesis. J Cell Biol 29 November 1999; 147 (5): 969–980. doi: https://doi.org/10.1083/jcb.147.5.969
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