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TitleHow Dedicated Ribosomes Translate a Leaderless mRNA.
Journal, issue, pagesJ Mol Biol, Vol. 436, Issue 4, Page 168423, Year 2024
Publish dateFeb 15, 2024
AuthorsFrancisco J Acosta-Reyes / Sayan Bhattacharjee / Max Gottesman / Joachim Frank /
PubMed AbstractIn bacteriophage λ lysogens, the λcI repressor is encoded by the leaderless transcript (lmRNA) initiated at the λpRM promoter. Translation is enhanced in rpsB mutants deficient in ribosomal ...In bacteriophage λ lysogens, the λcI repressor is encoded by the leaderless transcript (lmRNA) initiated at the λpRM promoter. Translation is enhanced in rpsB mutants deficient in ribosomal protein uS2. Although translation initiation of lmRNA is conserved in bacteria, archaea, and eukaryotes, structural insight of a lmRNA translation initiation complex is missing. Here, we use cryo-EM to solve the structures of the uS2-deficient 70S ribosome of host E. coli mutant rpsB11 and the wild-type 70S complex with λcI lmRNA and fMet-tRNA. Importantly, the uS2-deficient 70S ribosome also lacks protein bS21. The anti-Shine-Dalgarno (aSD) region is structurally supported by bS21, so that the absence of the latter causes the aSD to divert from the normal mRNA exit pathway, easing the exit of lmRNA. A π-stacking interaction between the monitor base A1493 and A(+4) of lmRNA potentially acts as a recognition signal. Coulomb charge flow, along with peristalsis-like dynamics within the mRNA entrance channel due to the increased 30S head rotation caused by the absence of uS2, are likely to facilitate the propagation of lmRNA through the ribosome. These findings lay the groundwork for future research on the mechanism of translation and the co-evolution of lmRNA and mRNA that includes the emergence of a defined ribosome-binding site of the transcript.
External linksJ Mol Biol / PubMed:38185325 / PubMed Central
MethodsEM (single particle)
Resolution3.2 - 3.3 Å
Structure data

41049

8t5d

EMDB-41049, PDB-8t5d:
Cryo-EM studies of the interplay between uS2 ribosomal protein and leaderless mRNA during bacterial translation initiation
Method: EM (single particle) / Resolution: 3.2 Å

41050

8t5h

EMDB-41050, PDB-8t5h:
Cryo-EM studies of the interplay between uS2 ribosomal protein and leaderless mRNA during bacterial translation initiation
Method: EM (single particle) / Resolution: 3.3 Å

Source
  • escherichia coli (E. coli)
  • bacteriophage sp. (virus)
KeywordsRIBOSOME / leaderless mRNA / Cryo-EM / 70S / HflX / bS21 / uS2

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