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Published online 25 December 2000. doi:10.1083/jcb.151.7.1561
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© The Rockefeller University Press, 0021-9525/2000//1561 $5.00
The Journal of Cell Biology, Volume 151, Number 7, , 2000 1561-1574

Original Article

In Vivo Analysis of Cajal Body Movement, Separation, and Joining in Live Human Cells



Melpomeni Platania, Ilya Goldbergb, Jason R. Swedlowa, and Angus I. Lamonda

a MSI/WTB Complex, University of Dundee, Dundee DD1 5EH, Scotland
b Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139
Department of Biochemistry, MSI/WTB Complex, University of Dundee, Dow St., Dundee DD1 5EH, Scotland, UK.44-1382-34569544-1382-345473

a.i.lamond{at}dundee.ac.uk

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.

Key Words: nucleus • Cajal bodies • coilin • GFP • time-lapse microscopy

© 2000 The Rockefeller University Press

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.


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