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ID: 176207.0, MPI für biophysikalische Chemie / Molekulare Biologie (Dr. Thomas M. Jovin)
RNAi knockdown of hPrp31 leads to an accumulation of U4/U6 di-snRNPs in Cajal bodies
Authors:Schaffert, N.; Hossbach, M.; Heintzmann, R.; Achsel, T.; Luehrmann, R.
Date of Publication (YYYY-MM-DD):2004-08-16
Title of Journal:EMBO Journal
Start Page:3000
End Page:3009
Sequence Number of Article:doi:10.1038/sj.emboj.7600296
Copyright:c* 2004 by EMBO

Jahrbuch 2004
Review Status:Peer-review
Audience:Not Specified
Abstract / Description:Cajal bodies (CBs) are subnuclear organelles of animal and plant cells. A role of CBs in the assembly and maturation of small nuclear ribonucleoproteins (snRNP) has been proposed but is poorly understood. Here we have addressed the question where U4/U6.U5 tri-snRNP assembly occurs in the nucleus. The U4/U6.U5 tri-snRNP is a central unit of the spliceosome and must be reformed from its components after each round of splicing. By combining RNAi and biochemical methods, we demonstrate that, after knockdown of the U4/U6-specific hPrp31 (61K) or the U5-specific hPrp6 (102 K) protein in HeLa cells, tri-snRNP formation is inhibited and stable U5 monosnRNPs and U4/U6 di-snRNPs containing U4/U6 proteins and the U4/U6 recycling factor p110 accumulate. Thus, hPrp31 and hPrp6 form an essential connection between the U4/U6 and U5 snRNPs in vivo. Using fluorescence microscopy, we show that, in the absence of either hPrp31 or hPrp6, U4/U6 di-snRNPs as well as p110 accumulate in Cajal bodies. In contrast, U5 snRNPs largely remain in nucleoplasmic speckles. Our data support the idea that CBs may play a role in tri-snRNP recycling.
Free Keywords:membranes & transport; RNA; Cajal bodies; hPrp31; pre-mRNA splicing; retinitis pigmentosa; snRNP biogenesis
Last Change of the Resource (YYYY-MM-DD):2004-08-16
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für biophysikalische Chemie/Abt. Reinhard Lührmann / 100
MPI für biophysikalische Chemie/Abt. Thomas Jovin / 060
External Affiliations:IRCCS Fondazione Santa Lucia, Neurobiologia, Via Ardeatina 306, 00179 Rome, Italy
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