Widespread Cotranslational Formation of Protein Complexes

Abstract

Most cellular processes are conducted by multi-protein complexes. However, little is known about how these complexes are assembled. In particular, it is not known if they are formed while one or more members of the complexes are being translated (cotranslational assembly). We took a genomic approach to address this question, by systematically identifying mRNAs associated with specific proteins. In a sample of 31 proteins from Schizosaccharomyces pombe that did not contain RNA–binding domains, we found that ∼38% copurify with mRNAs that encode interacting proteins. For example, the cyclin-dependent kinase Cdc2p associates with the rum1 and cdc18 mRNAs, which encode, respectively, an inhibitor of Cdc2p kinase activity and an essential regulator of DNA replication. Both proteins interact with Cdc2p and are key cell cycle regulators. We obtained analogous results with proteins with different structures and cellular functions (kinesins, protein kinases, transcription factors, proteasome components, etc.). We showed that copurification of a bait protein and of specific mRNAs was dependent on the presence of the proteins encoded by the interacting mRNAs and on polysomal integrity. These results indicate that these observed associations reflect the cotranslational interaction between the bait and the nascent proteins encoded by the interacting mRNAs. Therefore, we show that the cotranslational formation of protein–protein interactions is a widespread phenomenon.

Author Summary

Most proteins do not function in isolation. Instead, they associate with other proteins to form complexes. Little is known about the assembly of protein complexes within cells. One possibility is that proteins are completely synthesised before they bind to each other. An alternative is that proteins attach to each other as they are being translated in the ribosome (called cotranslational assembly). To investigate if cells use cotranslational assembly to form complexes, we identified mRNAs associated with specific proteins. The expectation is that if protein A binds to protein B as protein B is being translated, A will associate indirectly to the mRNA encoding B. Indeed, we found that for ∼40% of proteins (out of a sample of over 30) this was the case. Proteins associated with a small number of mRNAs, most of which encoded known or predicted interacting proteins. We found examples of this phenomenon in proteins with different functions and structures, indicating that cotranslational assembly is widespread. Cotranslational assembly might be required for certain proteins to associate, or it might be important in cases where the early formation of a protein complex is beneficial, such as when a protein is toxic or unstable unless bound to a partner.