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
作者： Cornelius Schneider , Dmitry E. Agafonov , Jana Schmitzová , Klaus Hartmuth , Patrizia Fabrizio
DOI: 10.1371/JOURNAL.PGEN.1005539
关键词: RNA splicing 、 Biology 、 Protein–protein interaction 、 Cell biology 、 Intron 、 Nucleotide 、 Precursor mRNA 、 Splicing factor 、 Spliceosome 、 Biochemistry 、 RNA Splicing Factors 、 Genetics(clinical) 、 Genetics 、 Cancer research 、 Ecology, Evolution, Behavior and Systematics 、 Molecular biology
摘要: Little is known about contacts in the spliceosome between proteins and intron nucleotides surrounding pre-mRNA branch-site their dynamics during splicing. We investigated protein-pre-mRNA interactions by UV-induced crosslinking of purified yeast Bact spliceosomes formed on site-specifically labeled pre-mRNA, analyzed changes after conversion to catalytically-activated B* step 1 C complexes, using a splicing system. Contacts upstream downstream U2 SF3a/b Prp9, Prp11, Hsh49, Cus1 Hsh155 were detected, demonstrating that these are evolutionarily conserved. The RES Pml1 Bud13 shown contact branch-site. A comparison pattern versus complexes revealed protein with dynamic. Upon catalysis, Cwc25 region, enhanced crosslinks Prp8 Prp45 observed. Cwc25’s promoting activity was not dependent its interaction indicating it acts via protein-protein interactions. These studies provide important insights into spliceosome's network reveal novel RNP remodeling events catalytic activation
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