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TitleRbfA Is Involved in Two Important Stages of 30S Subunit Assembly: Formation of the Central Pseudoknot and Docking of Helix 44 to the Decoding Center.
Journal, issue, pagesInt J Mol Sci, Vol. 22, Issue 11, Year 2021
Publish dateJun 7, 2021
AuthorsElena M Maksimova / Alexey P Korepanov / Olesya V Kravchenko / Timur N Baymukhametov / Alexander G Myasnikov / Konstantin S Vassilenko / Zhanna A Afonina / Elena A Stolboushkina /
PubMed AbstractRibosome biogenesis is a highly coordinated and complex process that requires numerous assembly factors that ensure prompt and flawless maturation of ribosomal subunits. Despite the increasing amount ...Ribosome biogenesis is a highly coordinated and complex process that requires numerous assembly factors that ensure prompt and flawless maturation of ribosomal subunits. Despite the increasing amount of data collected, the exact role of most assembly factors and mechanistic details of their operation remain unclear, mainly due to the shortage of high-resolution structural information. Here, using cryo-electron microscopy, we characterized 30S ribosomal particles isolated from an   strain with a deleted gene for the RbfA factor. The cryo-EM maps for pre-30S subunits were divided into six classes corresponding to consecutive assembly intermediates: from the particles with a completely unresolved head domain and unfolded central pseudoknot to almost mature 30S subunits with well-resolved body, platform, and head domains and partially distorted helix 44. The structures of two predominant 30S intermediates belonging to most populated classes obtained at 2.7 Å resolutions indicate that RbfA acts at two distinctive 30S assembly stages: early formation of the central pseudoknot including folding of the head, and positioning of helix 44 in the decoding center at a later stage. Additionally, it was shown that the formation of the central pseudoknot may promote stabilization of the head domain, likely through the RbfA-dependent maturation of the neck helix 28. An update to the model of factor-dependent 30S maturation is proposed, suggesting that RfbA is involved in most of the subunit assembly process.
External linksInt J Mol Sci / PubMed:34200244 / PubMed Central
MethodsEM (single particle)
Resolution2.69 - 12.0 Å
Structure data

EMDB-12854:
E.coli pre-30S delta rbfA ribosome subnit class E
Method: EM (single particle) / Resolution: 3.74 Å

EMDB-12855:
E. coli pre-30S delta rbfA ribosomal subunit class A
Method: EM (single particle) / Resolution: 12.0 Å

12856

7oe0

EMDB-12856, PDB-7oe0:
E. coli pre-30S delta rbfA ribosomal subunit class F
Method: EM (single particle) / Resolution: 2.69 Å

12857

7oe1

EMDB-12857, PDB-7oe1:
30S ribosomal subunit from E. coli
Method: EM (single particle) / Resolution: 3.05 Å

12916

7oi0

EMDB-12916, PDB-7oi0:
E.coli delta rbfA pre-30S ribosomal subunit class D
Method: EM (single particle) / Resolution: 2.76 Å

Chemicals

ChemComp-HOH:
WATER

Source
  • escherichia coli bw25113 (bacteria)
  • escherichia coli str. k-12 substr. mg1655 (bacteria)
  • escherichia coli (strain k12) (bacteria)
KeywordsRIBOSOME / 30S subunit / RNA / RbfA

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