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Published online 10 February 2003. doi:10.1083/jcb.200210087
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© The Rockefeller University Press, 0021-9525/2003/2/505 $5.00
The Journal of Cell Biology, Volume 160, Number 4, 505-516

Article

 Targeting of U4/U6 small nuclear RNP assembly factor SART3/p110 to Cajal bodies

David Stank1, Stephen D. Rader2, Mirko Klingauf1 and Karla M. Neugebauer1

1 Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany
2 Department of Biochemistry and Biophysics, University of California, San Francisco, CA, 94143

Address correspondence to Karla Neugebauer, Ph.D., Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany. Tel.: 49-351-2102589. Fax: 49-351-2101209. E-mail: neugebauer{at}mpi-cbg.de

The spliceosomal small nuclear RNAs (snRNAs) are distributed throughout the nucleoplasm and concentrated in nuclear inclusions termed Cajal bodies (CBs). A role for CBs in the metabolism of snRNPs has been proposed but is not well understood. The SART3/p110 protein interacts transiently with the U6 and U4/U6 snRNPs and promotes the reassembly of U4/U6 snRNPs after splicing in vitro. Here we report that SART3/p110 is enriched in CBs but not in gems or residual CBs lacking coilin. The U6 snRNP Sm-like (LSm) proteins, also involved in U4/U6 snRNP assembly, were localized to CBs as well. The levels of SART3/p110 and LSm proteins in CBs were reduced upon treatment with the transcription inhibitor {alpha}-amanitin, suggesting that CB localization reflects active processes dependent on transcription/splicing. The NH2-terminal HAT domain of SART3/p110 was necessary and sufficient for specific protein targeting to CBs. Overexpression of truncation mutants containing the HAT domain had dominant negative effects on U6 snRNP localization to CBs, indicating that endogenous SART3/p110 plays a role in targeting the U6 snRNP to CBs. We propose that U4 and U6 snRNPs accumulate in CBs for the purpose of assembly into U4/U6 snRNPs by SART3/p110.

Key Words: RNA splicing; coiled (Cajal) body; U4/U6 small nuclear ribonucleoprotein; small nuclear RNA; nuclear proteins


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