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Structure paper

TitleSequence-specific remodeling of a topologically complex RNP substrate by Spb4.
Journal, issue, pagesNat Struct Mol Biol, Vol. 29, Issue 12, Page 1228-1238, Year 2022
Publish dateDec 8, 2022
AuthorsVictor Emmanuel Cruz / Kamil Sekulski / Nagesh Peddada / Carolin Sailer / Sahana Balasubramanian / Christine S Weirich / Florian Stengel / Jan P Erzberger /
PubMed AbstractDEAD-box ATPases are ubiquitous enzymes essential in all aspects of RNA biology. However, the limited in vitro catalytic activities described for these enzymes are at odds with their complex cellular ...DEAD-box ATPases are ubiquitous enzymes essential in all aspects of RNA biology. However, the limited in vitro catalytic activities described for these enzymes are at odds with their complex cellular roles, most notably in driving large-scale RNA remodeling steps during the assembly of ribonucleoproteins (RNPs). We describe cryo-EM structures of 60S ribosomal biogenesis intermediates that reveal how context-specific RNA unwinding by the DEAD-box ATPase Spb4 results in extensive, sequence-specific remodeling of rRNA secondary structure. Multiple cis and trans interactions stabilize Spb4 in a post-catalytic, high-energy intermediate that drives the organization of the three-way junction at the base of rRNA domain IV. This mechanism explains how limited strand separation by DEAD-box ATPases is leveraged to provide non-equilibrium directionality and ensure efficient and accurate RNP assembly.
External linksNat Struct Mol Biol / PubMed:36482249 / PubMed Central
MethodsEM (single particle)
Resolution2.76 - 3.71 Å
Structure data

EMDB-24269: State E2 nucleolar 60S ribosomal biogenesis intermediate - Overall map
PDB-7nac: State E2 nucleolar 60S ribosomal biogenesis intermediate - Composite model
Method: EM (single particle) / Resolution: 3.04 Å

EMDB-24270, PDB-7nad:
State E2 nucleolar 60S ribosomal biogenesis intermediate - Spb4 local refinement model
Method: EM (single particle) / Resolution: 3.04 Å

EMDB-24271: State E2 nucleolar 60S ribosomal biogenesis intermediate - Spb1-MTD locally refined map
PDB-7naf: State E2 nucleolar 60S ribosomal biogenesis intermediate - Spb1-MTD local model
Method: EM (single particle) / Resolution: 3.13 Å

EMDB-24280: State E2 nucleolar 60S ribosomal intermediate - Local Map for Noc2/Noc3 region
PDB-7r6k: State E2 nucleolar 60S ribosomal intermediate - Model for Noc2/Noc3 region
Method: EM (single particle) / Resolution: 3.17 Å

EMDB-24286: State E2 nucleolar 60S ribosome biogenesis intermediate - Foot region map
PDB-7r6q: State E2 nucleolar 60S ribosome biogenesis intermediate - Foot region model
Method: EM (single particle) / Resolution: 2.98 Å

EMDB-24290: State E1 nucleolar 60S ribosome biogenesis intermediate - Spb4 locally refined map
PDB-7r72: State E1 nucleolar 60S ribosome biogenesis intermediate - Spb4 local model
Method: EM (single particle) / Resolution: 3.07 Å

EMDB-24296, PDB-7r7a:
State E1 nucleolar 60S ribosome biogenesis intermediate - Composite model
Method: EM (single particle) / Resolution: 3.04 Å

EMDB-24297: State E2 nucleolar 60S ribosomal biogenesis intermediate - L1 stalk local map
PDB-7r7c: State E2 nucleolar 60S ribosomal biogenesis intermediate - L1 stalk local model
Method: EM (single particle) / Resolution: 3.71 Å

EMDB-26259: State NE1 nucleolar 60S ribosome biogenesis intermediate - Overall map
PDB-7u0h: State NE1 nucleolar 60S ribosome biogenesis intermediate - Overall model
Method: EM (single particle) / Resolution: 2.76 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-SAH:
S-ADENOSYL-L-HOMOCYSTEINE

ChemComp-HOH:
WATER

ChemComp-MG:
Unknown entry

Source
  • saccharomyces cerevisiae by4741 (yeast)
KeywordsRIBOSOME / ribosome biogenesis / DEAD-box ATPases / methyltransferase / nucleolus / methyltransferases / DEAD-box ATPase

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