Journal article

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


Authors listCross, M; Günzl, A; Palfi, Z; Bindereif, A

Publication year1991

Pages5516-5526

JournalMolecular and Cellular Biology

Volume number11

Issue number11

ISSN0270-7306

eISSN1098-5549

DOI Linkhttps://doi.org/10.1128/MCB.11.11.5516

PublisherTaylor and Francis Group


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.



Citation Styles

Harvard Citation styleCross, M., Günzl, A., Palfi, Z. and Bindereif, A. (1991) Analysis of small nuclear ribonucleoproteins (RNPs) in Trypanosoma brucei: structural organization and protein components of the spliced leader RNP., Molecular and Cellular Biology, 11(11), pp. 5516-5526. https://doi.org/10.1128/MCB.11.11.5516

APA Citation styleCross, M., Günzl, A., Palfi, Z., & Bindereif, A. (1991). Analysis of small nuclear ribonucleoproteins (RNPs) in Trypanosoma brucei: structural organization and protein components of the spliced leader RNP.. Molecular and Cellular Biology. 11(11), 5516-5526. https://doi.org/10.1128/MCB.11.11.5516


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