Dynamic Contacts of U2, RES, Cwc25, Prp8 and Prp45 Proteins with the Pre-mRNA Branch-Site and 3' Splice Site during Catalytic Activation and Step 1 Catalysis in Yeast Spliceosomes


The spliceosome is a highly dynamic RNP machine that during the catalytic cycle undergoes many changes in composition and conformation. The pre-catalytic Bact spliceosome contains the U2, U6 and U5 snRNAs and ~40 proteins, which are evolutionarily conserved between budding yeast and metazoans. The Bact spliceosome is converted to a catalytically-activated B* spliceosome and following recruitment of the Cwc25 protein, step 1 of splicing is catalyzed and the C spliceosome is generated. The U2 snRNP plays an essential role in branch-site selection and pre-mRNA splicing catalysis. During the Bact to B* transition the affinity of several U2 SF3a/b proteins for the spliceosome is significantly reduced. Whether this is due to remodeling events affecting U2 snRNP contacts with the pre-mRNA is not known. Information about conserved spliceosomal protein-pre-mRNA contacts and their dynamics during splicing remains limited. Here we investigated pre-mRNA–protein contact sites in yeast Bact spliceosomes by UV-induced crosslinking. We detected contacts of nucleotides surrounding the branch-site with several of the U2 SF3a/b proteins, and we show that these interactions are evolutionarily conserved. We carried out a similar investigation with B* and C spliceosomes and provide important insights into the dynamics of pre-mRNA–protein interactions involving the essential U2, RES, Cwc25, Prp8 and Prp45 proteins.


Vyšlo v časopise: Dynamic Contacts of U2, RES, Cwc25, Prp8 and Prp45 Proteins with the Pre-mRNA Branch-Site and 3' Splice Site during Catalytic Activation and Step 1 Catalysis in Yeast Spliceosomes. PLoS Genet 11(9): e32767. doi:10.1371/journal.pgen.1005539
Kategorie: Research Article
prolekare.web.journal.doi_sk: 10.1371/journal.pgen.1005539

Souhrn

The spliceosome is a highly dynamic RNP machine that during the catalytic cycle undergoes many changes in composition and conformation. The pre-catalytic Bact spliceosome contains the U2, U6 and U5 snRNAs and ~40 proteins, which are evolutionarily conserved between budding yeast and metazoans. The Bact spliceosome is converted to a catalytically-activated B* spliceosome and following recruitment of the Cwc25 protein, step 1 of splicing is catalyzed and the C spliceosome is generated. The U2 snRNP plays an essential role in branch-site selection and pre-mRNA splicing catalysis. During the Bact to B* transition the affinity of several U2 SF3a/b proteins for the spliceosome is significantly reduced. Whether this is due to remodeling events affecting U2 snRNP contacts with the pre-mRNA is not known. Information about conserved spliceosomal protein-pre-mRNA contacts and their dynamics during splicing remains limited. Here we investigated pre-mRNA–protein contact sites in yeast Bact spliceosomes by UV-induced crosslinking. We detected contacts of nucleotides surrounding the branch-site with several of the U2 SF3a/b proteins, and we show that these interactions are evolutionarily conserved. We carried out a similar investigation with B* and C spliceosomes and provide important insights into the dynamics of pre-mRNA–protein interactions involving the essential U2, RES, Cwc25, Prp8 and Prp45 proteins.


Zdroje

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