title: Dynamic protein-protein interaction wiring of the human spliceosome.
abstract: More than 200 proteins copurify with spliceosomes, the compositionally dynamic RNPs catalyzing pre-mRNA splicing. To better understand protein - protein interactions governing splicing, we systematically investigated interactions between human spliceosomal proteins. A comprehensive Y2H interaction matrix screen generated a protein interaction map comprising 632 interactions between 196 proteins. Among these, 242 interactions were found between spliceosomal core proteins and largely validated by coimmunoprecipitation. To reveal dynamic changes in protein interactions, we integrated spliceosomal complex purification information with our interaction data and performed link clustering. These data, together with interaction competition experiments, suggest that during step 1 of splicing, hPRP8 interactions with SF3b proteins are replaced by hSLU7, positioning this second step factor close to the active site, and that the DEAH-box helicases hPRP2 and hPRP16 cooperate through ordered interactions with GPKOW. Our data provide extensive information about the spliceosomal protein interaction network and its dynamics.
authors.affiliation: Otto-Warburg Laboratory, Max-Planck Institute for Molecular Genetics, 14195 Berlin, Germany.
authors.forename: Anna
authors.initials: A
authors.lastname: Hegele
authors.forename: Atanas
authors.initials: A
authors.lastname: Kamburov
authors.forename: Arndt
authors.initials: A
authors.lastname: Grossmann
authors.forename: Chrysovalantis
authors.initials: C
authors.lastname: Sourlis
authors.forename: Sylvia
authors.initials: S
authors.lastname: Wowro
authors.forename: Mareike
authors.initials: M
authors.lastname: Weimann
authors.forename: Cindy L
authors.initials: CL
authors.lastname: Will
authors.forename: Vlad
authors.initials: V
authors.lastname: Pena
authors.forename: Reinhard
authors.initials: R
authors.lastname: Lührmann
authors.forename: Ulrich
authors.initials: U
authors.lastname: Stelzl
mesh_terms: D001667:Binding, Competitive; D002352:Carrier Proteins / Q000378:metabolism; D016000:Cluster Analysis; D053487:DEAD-box RNA Helicases / Q000378:metabolism / Q000502:physiology; D006801:Humans; D054730:Protein Interaction Domains and Motifs; D060066:Protein Interaction Maps; D040901:Proteomics; D020365:RNA Helicases / Q000378:metabolism / Q000502:physiology; D012322:RNA Precursors / Q000378:metabolism*; D012326:RNA Splicing; D000072260:RNA Splicing Factors; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism / Q000502:physiology; D017411:Ribonucleoproteins, Small Nuclear / Q000378:metabolism; D017381:Spliceosomes / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; C115374:PRPF8 protein, human; D012322:RNA Precursors; D000072260:RNA Splicing Factors; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D017411:Ribonucleoproteins, Small Nuclear; C490656:SLU7 protein, human; C555647:DHX16 protein, human; C112199:DHX38 protein, human; D053487:DEAD-box RNA Helicases; D020365:RNA Helicases