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Structure paper

TitleMolecular basis of mRNA delivery to the bacterial ribosome.
Journal, issue, pagesScience, Vol. 386, Issue 6725, Page eado8476, Year 2024
Publish dateNov 29, 2024
AuthorsMichael W Webster / Adrien Chauvier / Huma Rahil / Andrea Graziadei / Kristine Charles / Nataliya Miropolskaya / Maria Takacs / Charlotte Saint-André / Juri Rappsilber / Nils G Walter / Albert Weixlbaumer /
PubMed AbstractProtein synthesis begins with the formation of a ribosome-messenger RNA (mRNA) complex. In bacteria, the small ribosomal subunit (30) is recruited to many mRNAs through base pairing with the Shine- ...Protein synthesis begins with the formation of a ribosome-messenger RNA (mRNA) complex. In bacteria, the small ribosomal subunit (30) is recruited to many mRNAs through base pairing with the Shine-Dalgarno (SD) sequence and RNA binding by ribosomal protein bS1. Translation can initiate on nascent mRNAs, and RNA polymerase (RNAP) can promote the recruitment of the pioneering 30. Here, we examined 30 recruitment to nascent mRNAs using cryo-electron microscopy, single-molecule fluorescence colocalization, and in-cell cross-linking mass spectrometry. We show that bS1 delivers the mRNA to the ribosome for SD duplex formation and 30 activation. Additionally, bS1 and RNAP stimulate translation initiation. Our work provides a mechanistic framework for how the SD duplex, ribosomal proteins, and RNAP cooperate in 30 recruitment to mRNAs and establish transcription-translation coupling.
External linksScience / PubMed:39607923
MethodsEM (single particle)
Resolution2.5 - 4.2 Å
Structure data

51517

9gr1

EMDB-51517, PDB-9gr1:
E. coli 70S-TEC complex in delivery state
Method: EM (single particle) / Resolution: 3.17 Å

EMDB-51572: 30S mRNA delivery complex (open head) consensus map
Method: EM (single particle) / Resolution: 3.0 Å

EMDB-51573: 30S mRNA delivery complex (open head) focused head map
Method: EM (single particle) / Resolution: 3.0 Å

EMDB-51574: 30S mRNA delivery complex (open head) focused body map
Method: EM (single particle) / Resolution: 3.0 Å

EMDB-51575: 30S mRNA delivery complex (open head) focused platform map
Method: EM (single particle) / Resolution: 3.0 Å

EMDB-51576: 30S mRNA delivery complex (consensus) consensus map
Method: EM (single particle) / Resolution: 2.6 Å

EMDB-51577: 30S mRNA delivery complex (consensus) focused head map
Method: EM (single particle) / Resolution: 2.5 Å

EMDB-51578: 30S mRNA delivery complex (consensus) focused body map
Method: EM (single particle) / Resolution: 2.5 Å

EMDB-51579: 30S mRNA delivery complex (consensus) focused platform map
Method: EM (single particle) / Resolution: 2.5 Å

EMDB-51580: 30S mRNA delivery complex (bS1 resolved) consensus map
Method: EM (single particle) / Resolution: 2.9 Å

EMDB-51581: 30S mRNA delivery complex (bS1 resolved) focused head map
Method: EM (single particle) / Resolution: 2.8 Å

EMDB-51582: 30S mRNA delivery complex (bS1 resolved) focused body map
Method: EM (single particle) / Resolution: 2.8 Å

EMDB-51583: 30S mRNA delivery complex (bS1 resolved) focused platform map
Method: EM (single particle) / Resolution: 2.8 Å

EMDB-51584: 30S mRNA delivery complex TEC resolved (30S only) consensus map
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-51585: 30S mRNA delivery complex TEC resolved (30S only) focused head map
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-51586: 30S mRNA delivery complex TEC resolved (30S only) focused body map
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-51587: 30S mRNA delivery complex (closed head) consensus map
Method: EM (single particle) / Resolution: 2.7 Å

EMDB-51588: 30S mRNA delivery complex (closed head) focused head map
Method: EM (single particle) / Resolution: 2.6 Å

EMDB-51589: 30S mRNA delivery complex (closed head) focused body map
Method: EM (single particle) / Resolution: 2.7 Å

EMDB-51590: 30S mRNA delivery complex (closed head) focused platform map
Method: EM (single particle) / Resolution: 2.6 Å

EMDB-51591: 30S-TEC (TEC in expressome position) Inactive state 2, consensus map
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-51592: 30S-TEC (TEC in expressome position) Inactive state 2, focused head map
Method: EM (single particle) / Resolution: 3.0 Å

EMDB-51593: 30S-TEC (TEC in expressome position) Inactive state 2, focused body map
Method: EM (single particle) / Resolution: 3.0 Å

EMDB-51594: 30S-TEC (TEC in expressome position) Inactive state 2, focused platform map
Method: EM (single particle) / Resolution: 3.0 Å

EMDB-51595: 30S-TEC (TEC in expressome position) Inactive state 2, focused TEC map
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-51596: 30S-TEC (TEC in expressome position) Inactive state 1, consensus map
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-51597: 30S-TEC (TEC in expressome position) Inactive state 1, focused head map
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-51598: 30S-TEC (TEC in expressome position) Inactive state 1, focused body map
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-51599: 30S-TEC (TEC in expressome position) Inactive state 1, focused platform map
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-51600: 30S-TEC (TEC in expressome position) Inactive state 1, focused TEC map
Method: EM (single particle) / Resolution: 3.4 Å

EMDB-51601: 30S mRNA delivery complex TEC resolved (30S only) focused platform map
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-51602: 30S-PIC (Pre-Initiation complex) consensus map
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-51603: 30S-PIC (Pre-Initiation complex) focused head map
Method: EM (single particle) / Resolution: 3.1 Å

EMDB-51604: 30S-PIC (Pre-Initiation complex) focused body map
Method: EM (single particle) / Resolution: 3.1 Å

51615

9gup

EMDB-51615, PDB-9gup:
30S mRNA delivery complex (open head)
Method: EM (single particle) / Resolution: 2.8 Å

51616

9guq

EMDB-51616, PDB-9guq:
30S PIC (Pre-Initiation complex)
Method: EM (single particle) / Resolution: 3.1 Å

51617

9gur

EMDB-51617, PDB-9gur:
30S mRNA delivery complex TEC resolved (TEC only)
Method: EM (single particle) / Resolution: 4.2 Å

51618

9gus

EMDB-51618, PDB-9gus:
30S mRNA delivery complex TEC resolved (30S only)
Method: EM (single particle) / Resolution: 3.5 Å

51619

9gut

EMDB-51619, PDB-9gut:
30S mRNA delivery complex (bS1 resolved)
Method: EM (single particle) / Resolution: 2.8 Å

51620

9guu

EMDB-51620, PDB-9guu:
30S mRNA delivery complex (consensus)
Method: EM (single particle) / Resolution: 2.5 Å

51621

9guv

EMDB-51621, PDB-9guv:
30S mRNA delivery complex (closed-head)
Method: EM (single particle) / Resolution: 3.0 Å

51622

9guw

EMDB-51622, PDB-9guw:
30S-TEC (TEC in expressome position) Inactive state 2
Method: EM (single particle) / Resolution: 3.1 Å

51623

9gux

EMDB-51623, PDB-9gux:
30S-TEC (TEC in expressome position) Inactive state 1
Method: EM (single particle) / Resolution: 3.3 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-K:
Unknown entry

ChemComp-MG:
Unknown entry

ChemComp-PHE:
PHENYLALANINE

Source
  • escherichia coli bl21(de3) (bacteria)
  • synthetic construct (others)
  • escherichia coli (E. coli)
  • escherichia coli k-12 (bacteria)
KeywordsGENE REGULATION / Ribosome / RNA polymerase / mRNA / Transcription / translation / coupling / Ribosome. / coupling.

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