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TitleCryo-EM reveals active site coordination within a multienzyme pre-rRNA processing complex.
Journal, issue, pagesNat. Struct. Mol. Biol., Vol. 26, Issue 9, Page 830-839, Year 2019
Publish dateSep 5, 2019
AuthorsMonica C Pillon / Allen L Hsu / Juno M Krahn / Jason G Williams / Kevin H Goslen / Mack Sobhany / Mario J Borgnia / Robin E Stanley /
PubMed AbstractRibosome assembly is a complex process reliant on the coordination of trans-acting enzymes to produce functional ribosomal subunits and secure the translational capacity of cells. The ...Ribosome assembly is a complex process reliant on the coordination of trans-acting enzymes to produce functional ribosomal subunits and secure the translational capacity of cells. The endoribonuclease (RNase) Las1 and the polynucleotide kinase (PNK) Grc3 assemble into a multienzyme complex, herein designated RNase PNK, to orchestrate processing of precursor ribosomal RNA (rRNA). RNase PNK belongs to the functionally diverse HEPN nuclease superfamily, whose members rely on distinct cues for nuclease activation. To establish how RNase PNK coordinates its dual enzymatic activities, we solved a series of cryo-EM structures of Chaetomium thermophilum RNase PNK in multiple conformational states. The structures reveal that RNase PNK adopts a butterfly-like architecture, harboring a composite HEPN nuclease active site flanked by discrete RNA kinase sites. We identify two molecular switches that coordinate nuclease and kinase function. Together, our structures and corresponding functional studies establish a new mechanism of HEPN nuclease activation essential for ribosome production.
External linksPubMed:31488907 / Publisher's page
KeywordsRNA BINDING PROTEIN / Complex / Ribonuclease / Polynucleotide kinase
MethodsEM (single particle)
Resolution2.9 - 3.2 A
Structure data

EMDB-20040:
Precursor ribosomal RNA processing complex, State 2.

EMDB-20041:
Precursor ribosomal RNA processing complex, State 1.

EMDB-20042:
Precursor ribosomal RNA processing complex, apo-state.

PDB-6of2:
Precursor ribosomal RNA processing complex, State 2.

PDB-6of3:
Precursor ribosomal RNA processing complex, State 1.

PDB-6of4:
Precursor ribosomal RNA processing complex, apo-state.

Chemicals

ChemComp-AGS:
PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER

ChemComp-MG:
MAGNESIUM IONMagnesium

Source
  • Chaetomium thermophilum (fungus)
  • chaetomium thermophilum (strain dsm 1495 / cbs 144.50 / imi 039719) (fungus)

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    • Jacques Dubochet (University of Lausanne, Switzerland) is a pioneer of ice-embedding method of EM specimen (as known as cryo-EM), Most of 3DEM structures in EMDB and PDB are obtained using his method.
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