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TitleThe CryoEM structure of the ribosome maturation factor Rea1.
Journal, issue, pagesElife, Vol. 7, Year 2018
Publish dateNov 26, 2018
AuthorsPiotr Sosnowski / Linas Urnavicius / Andreas Boland / Robert Fagiewicz / Johan Busselez / Gabor Papai / Helgo Schmidt /
PubMed AbstractThe biogenesis of 60S ribosomal subunits is initiated in the nucleus where rRNAs and proteins form pre-60S particles. These pre-60S particles mature by transiently interacting with various assembly ...The biogenesis of 60S ribosomal subunits is initiated in the nucleus where rRNAs and proteins form pre-60S particles. These pre-60S particles mature by transiently interacting with various assembly factors. The ~5000 amino-acid AAA+ ATPase Rea1 (or Midasin) generates force to mechanically remove assembly factors from pre-60S particles, which promotes their export to the cytosol. Here we present three Rea1 cryoEM structures. We visualise the Rea1 engine, a hexameric ring of AAA+ domains, and identify an α-helical bundle of AAA2 as a major ATPase activity regulator. The α-helical bundle interferes with nucleotide-induced conformational changes that create a docking site for the substrate binding MIDAS domain on the AAA +ring. Furthermore, we reveal the architecture of the Rea1 linker, which is involved in force generation and extends from the AAA+ ring. The data presented here provide insights into the mechanism of one of the most complex ribosome maturation factors.
External linksElife / PubMed:30460895 / PubMed Central
MethodsEM (single particle)
Resolution3.9 - 7.8 Å
Structure data

0308

6hyd

EMDB-0308, PDB-6hyd:
Rea1 Wild type ADP state (tail part)
Method: EM (single particle) / Resolution: 3.9 Å

0309

6hyp

EMDB-0309, PDB-6hyp:
Rea1 Wild type ADP state (AAA+ ring part)
Method: EM (single particle) / Resolution: 4.4 Å

0328

6i26

EMDB-0328, PDB-6i26:
Rea1 Wild type AMPPNP state
Method: EM (single particle) / Resolution: 4.3 Å

0329

6i27

EMDB-0329, PDB-6i27:
Rea1 AAA2L-H2alpha deletion mutant in AMPPNP State
Method: EM (single particle) / Resolution: 7.8 Å

EMDB-0330:
Rea1 Wild type ADP state (AAA+ ring + stem)
Method: EM (single particle) / Resolution: 4.4 Å

Chemicals

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM / Adenosine diphosphate

Source
  • Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast)
  • saccharomyces cerevisiae (brewer's yeast)
KeywordsMOTOR PROTEIN / Rea1 / Mdn1 / Midasin / AAA+ protein / ribosome maturation / molecular machine

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