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Published 11 October 2004. doi:10.1083/jcb.200404120
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 167, Number 1, 51-63
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Article

 Hypophosphorylated SR splicing factors transiently localize around active nucleolar organizing regions in telophase daughter nuclei

Paula A. Bubulya1, Kannanganattu V. Prasanth1, Thomas J. Deerinck2, Daniel Gerlich3, Joel Beaudouin3, Mark H. Ellisman2, Jan Ellenberg3, and David L. Spector1

1 Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724
2 National Center for Microscopy and Imaging Research, University of California, San Diego, La Jolla, CA 92093
3 Gene Expression and Cell Biology/Biophysics Programmes, European Molecular Biology Laboratory, Heidelberg D-69117 Germany

Correspondence to D.L. Spector: spector{at}cshl.org

Upon completion of mitosis, daughter nuclei assemble all of the organelles necessary for the implementation of nuclear functions. We found that upon entry into daughter nuclei, snRNPs and SR proteins do not immediately colocalize in nuclear speckles. SR proteins accumulated in patches around active nucleolar organizing regions (NORs) that we refer to as NOR-associated patches (NAPs), whereas snRNPs were enriched at other nuclear regions. NAPs formed transiently, persisting for 15–20 min before dissipating as nuclear speckles began to form in G1. In the absence of RNA polymerase II transcription, NAPs increased in size and persisted for at least 2 h, with delayed localization of SR proteins to nuclear speckles. In addition, SR proteins in NAPs are hypophosphorylated, and the SR protein kinase Clk/STY colocalizes with SR proteins in NAPs, suggesting that phosphorylation releases SR proteins from NAPs and their initial target is transcription sites. This work demonstrates a previously unrecognized role of NAPs in splicing factor trafficking and nuclear speckle biogenesis.

Abbreviations used in this paper: CB, Cajal body; IBB, importin-ß binding domain; IGC, interchromatin granule cluster; MIG, mitotic IGC; NAP, NOR-associated patch; NOR, nucleolar organizing region; PNC, perinucleolar compartment; PTB, polypyrimidine tract-binding protein; RS, arginine-serine rich.


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