This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Cross, M Articles by Bindereif, A Search for Related Content PubMed PubMed Citation Articles by Cross, M Articles by Bindereif, A

 Previous Article  |  Next Article 

Mol Cell Biol. 1991 November; 11(11): 5516-5526

Analysis of small nuclear ribonucleoproteins (RNPs) in Trypanosoma brucei: structural organization and protein components of the spliced leader RNP.

Max-Planck-Institut für Molekulare Genetik, Otto-Warburg-Laboratorium, Berlin, Dahlem, Germany.

ABSTRACT

trans splicing in Trypanosoma brucei involves the ligation of the 40-nucleotide spliced leader (SL) to each of the exons of large, polycistronic pre-mRNAs and requires the function of small nuclear ribonucleoproteins (snRNPs). We have identified and characterized snRNP complexes of SL, U2, U4, and U6 RNAs in T. brucei extracts by a combination of glycerol gradient sedimentation, CsCl density centrifugation, and anti-m3G immunoprecipitation. Both the SL RNP and the U4/U6 snRNP contain salt-stable cores; the U2 snRNP, in contrast to other eucaryotic snRNPs, is not stable under stringent ionic conditions. Two distinct complexes of U6 RNA were found, a U6 snRNP and a U4/U6 snRNP. The structure of the SL RNP was analyzed in detail by oligonucleotide-directed RNase H protection and by in vitro reconstitution. Our results indicate that the 3' half of SL RNA constitutes the core protein-binding domain and that protein components of the SL RNP also bind to the U2 and U4 RNAs. Using antisense RNA affinity chromatography, we identified a set of low-molecular-mass proteins (14.8, 14, 12.5, and 10 kDa) as components of the core SL RNP.

This article has been cited by other articles:

• Palfi, Z., Schimanski, B., Gunzl, A., Lucke, S., Bindereif, A. (2005). U1 small nuclear RNP from Trypanosoma brucei: a minimal U1 snRNA with unusual protein components. Nucleic Acids Res 33: 2493-2503 [Abstract] [Full Text]  
• Schnare, M. N., Gray, M. W. (1999). A Candidate U1 Small Nuclear RNA for Trypanosomatid Protozoa. J. Biol. Chem. 274: 23691-23694 [Abstract] [Full Text]  
• Goncharov, I., Palfi, Z., Bindereif, A., Michaeli, S. (1999). Purification of the Spliced Leader Ribonucleoprotein Particle from Leptomonas collosoma Revealed the Existence of an Sm Protein in Trypanosomes. CLONING THE SmE HOMOLOGUE. J. Biol. Chem. 274: 12217-12221 [Abstract] [Full Text]  
• Xu, Y.-x., Ben-Shlomo, H., Michaeli, S. (1997). The U5 RNA of trypanosomes deviates from the canonical U5 RNA: The Leptomonas collosoma U5 RNA and its coding gene. Proc. Natl. Acad. Sci. USA 94: 8473-8478 [Abstract] [Full Text]  
• Hinz, M., Moore, M. J., Bindereif, A. (1996). Domain Analysis of Human U5 RNA. CAP TRIMETHYLATION, PROTEIN BINDING, AND SPLICEOSOME ASSEMBLY. J. Biol. Chem. 271: 19001-19007 [Abstract] [Full Text]  
• Ullu, E., Tschudi, C. (1995). Accurate Modification of the Trypanosome Spliced Leader Cap Structure in a Homologous Cell-free System. J. Biol. Chem. 270: 20365-20369 [Abstract] [Full Text]  
• Wolff, T, Bindereif, A (1993). Conformational changes of U6 RNA during the spliceosome cycle: an intramolecular helix is essential both for initiating the U4-U6 interaction and for the first step of slicing.. Genes Dev. 7: 1377-1389 [Abstract]  
• Palfi, Z., Lucke, S., Lahm, H.-W., Lane, W. S., Kruft, V., Bragado-Nilsson, E., Seraphin, B., Bindereif, A. (2000). The spliceosomal snRNP core complex of Trypanosoma brucei: Cloning and functional analysis reveals seven Sm protein constituents. Proc. Natl. Acad. Sci. USA 97: 8967-8972 [Abstract] [Full Text]