Cajal bodies (also known as coiled bodies) are subnuclear organelles that contain specific nuclear antigens, including splicing small nuclear ribonucleoproteins (snRNPs) and a subset of nucleolar proteins. Cajal bodies are localized in the nucleoplasm and are often found at the nucleolar periphery. We have constructed a stable HeLa cell line, HeLaGFP-coilin, that expresses the Cajal body marker protein, p80 coilin, fused to the green fluorescent protein (GFP-coilin). The localization pattern and biochemical properties of the GFP-coilin fusion protein are identical to the endogenous p80 coilin. Time-lapse recordings on 63 nuclei of HeLaGFP-coilin cells showed that all Cajal bodies move within the nucleoplasm. Movements included translocations through the nucleoplasm, joining of bodies to form larger structures, and separation of smaller bodies from larger Cajal bodies. Also, we observed Cajal bodies moving to and from nucleoli. The data suggest that there may be at least two classes of Cajal bodies that differ in their size, antigen composition, and dynamic behavior. The smaller size class shows more frequent and faster rates of movement, up to 0.9 μm/min. The GFP-coilin protein is dynamically associated with Cajal bodies as shown by changes in their fluorescence intensity over time. This study reveals an unexpectedly high level of movement and interactions of nuclear bodies in human cells and suggests that these movements may be driven, at least in part, by regulated mechanisms.
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25 December 2000
Article|
December 25 2000
In Vivo Analysis of Cajal Body Movement, Separation, and Joining in Live Human Cells
Melpomeni Platani,
Melpomeni Platani
aMSI/WTB Complex, University of Dundee, Dundee DD1 5EH, Scotland
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Ilya Goldberg,
Ilya Goldberg
bDepartment of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139
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Jason R. Swedlow,
Jason R. Swedlow
aMSI/WTB Complex, University of Dundee, Dundee DD1 5EH, Scotland
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Angus I. Lamond
Angus I. Lamond
aMSI/WTB Complex, University of Dundee, Dundee DD1 5EH, Scotland
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Melpomeni Platani
aMSI/WTB Complex, University of Dundee, Dundee DD1 5EH, Scotland
Ilya Goldberg
bDepartment of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139
Jason R. Swedlow
aMSI/WTB Complex, University of Dundee, Dundee DD1 5EH, Scotland
Angus I. Lamond
aMSI/WTB Complex, University of Dundee, Dundee DD1 5EH, Scotland
The online version of this article contains supplemental material.
Abbreviations used in this paper: DIC, differential interference contrast; SIP, SMN-interacting protein; SMN, survival of motor neuron; sn, small nuclear; sno, small nucleolar; YFP, yellow fluorescent protein.
Received:
May 30 2000
Revision Requested:
October 16 2000
Accepted:
October 17 2000
Online ISSN: 1540-8140
Print ISSN: 0021-9525
© 2000 The Rockefeller University Press
2000
The Rockefeller University Press
J Cell Biol (2000) 151 (7): 1561–1574.
Article history
Received:
May 30 2000
Revision Requested:
October 16 2000
Accepted:
October 17 2000
Citation
Melpomeni Platani, Ilya Goldberg, Jason R. Swedlow, Angus I. Lamond; In Vivo Analysis of Cajal Body Movement, Separation, and Joining in Live Human Cells. J Cell Biol 25 December 2000; 151 (7): 1561–1574. doi: https://doi.org/10.1083/jcb.151.7.1561
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