301. Genome-Wide Protein Interaction Screens Reveal Functional Networks Involving Sm-Like Proteins
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Pierre Legrain, Laurence Decourty, Nicolas Joly, Florence Ricard, Jean D. Beggs, Andrew E. Mayes, Jean-Christophe Rain, Micheline Fromont-Racine, Ian Dix, Adeline Brunet-Simon, Alan Colley, Génétique des Intéractions Macromoléculaires, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), University of Edinburgh, Informatique scientifique, Institut Pasteur [Paris], This work was supported in part by the European Union (Biotech 95007—the TAPIR network), by a grant from GIP-HMR and by grant 047685 to JDB from the Wellcome Trust. JDB is supported by a Royal Society Cephalosporin Fund Senior Research Fellowship and AEM was recipient of a Wellcome Trust Prize Studentship., Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)
- Subjects
Proteome ,Article Subject ,RNA Splicing ,proteome ,snRNP ,[SDV]Life Sciences [q-bio] ,Saccharomyces cerevisiae ,Prp24 ,Bioengineering ,Computational biology ,yeast ,MESH: Two-Hybrid System Techniques ,Genome ,Applied Microbiology and Biotechnology ,Biochemistry ,Fungal Proteins ,splicing ,03 medical and health sciences ,Two-Hybrid System Techniques ,Genetics ,RNA, Messenger ,two-hybrid ,MESH: RNA, Messenger ,030304 developmental biology ,Fungal protein ,0303 health sciences ,biology ,MESH: RNA, Fungal ,MESH: Ribonucleoproteins, Small Nuclear ,030302 biochemistry & molecular biology ,RNA ,RNA, Fungal ,biology.organism_classification ,Ribonucleoproteins, Small Nuclear ,MESH: Saccharomyces cerevisiae ,MESH: Proteome ,mRNA degradation ,RNA splicing ,MESH: Genome, Fungal ,MESH: Fungal Proteins ,Genome, Fungal ,MESH: RNA Splicing ,Research Article ,Biotechnology - Abstract
A set of seven structurally related Sm proteins forms the core of the snRNP particles containing the spliceosomal U1, U2, U4 and U5 snRNAs. A search of the genomic sequence of Saccharomyces cerevisiae has identified a number of open reading frames that potentially encode structurally similar proteins termed Lsm (L¯ike Sm¯) proteins. With the aim of analysing all possible interactions between the Lsm proteins and any protein encoded in the yeast genome, we performed exhaustive and iterative genomic two-hybrid screens, starting with the Lsm proteins as baits. Indeed, extensive interactions amongst eight Lsm proteins were found that suggest the existence of a Lsm complex or complexes. These Lsm interactions apparently involve the conserved Sm domain that also mediates interactions between the Sm proteins. The screens also reveal functionally significant interactions with splicing factors, in particular with Prp4 and Prp24, compatible with genetic studies and with the reported association of Lsm proteins with spliceosomal U6 and U4/U6 particles. In addition, interactions with proteins involved in mRNA turnover, such as Mrt1, Dcp1, Dcp2 and Xrn1, point to roles for Lsm complexes in distinct RNA metabolic processes, that are confirmed in independent functional studies. These results provide compelling evidence that two-hybrid screens yield functionally meaningful information about protein–protein interactions and can suggest functions for uncharacterized proteins, especially when they are performed on a genome-wide scale.
- Published
- 2000
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