Recent X-ray crystallographic and cryo-electron microscopy data indicate that structured mRNAs bind to the bacterial ribosome at an early step of translation initiation, to a newly characterised ribosomal site located at the “platform” of the small ribosomal subunit (the “platform-binding” site or PBS). In a second step, the mRNAs are supposed to unfold and to migrate to the P-site to interact with the initiator tRNA. The proposal aims at dissecting the role of ribosomal proteins situated at the PBS and potentially contacting the structured 5’-terminal parts of mRNAs. It is proposed to isolate mutants in the genes of the corresponding proteins that influence the in vivo translation of the lacZ gene fused to different model 5’-UTRs known to be involved with translation regulation. Once such mutations are found, global analysis of their effect will be performed and their effect will be analyzed in vitro.
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