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PreprintCited by 13RNA and protein synthesis mechanismsNuclear Structure and FunctionRNA Research and Splicing

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Concurrent remodelling of nucleolar 60S subunit precursors by the Rea1 ATPase and Spb4 RNA helicase

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Mitterer, Valentin, Thoms, Matthias, Buschauer, Robert, Berninghausen, Otto, Hurt, Ed, Beckmann, Roland·2022·eLife
DOI 10.1101/2022.11.14.516466PMC PMC10154028

Preprint — published as 10.7554/elife.84877 · Paper page

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Abstract

Biogenesis intermediates of nucleolar ribosomal 60S precursor particles undergo a number of structural maturation steps before they transit to the nucleoplasm and are finally exported into the cytoplasm. The AAA+-ATPase Rea1 participates in the nucleolar exit by releasing the Ytm1–Erb1 heterodimer from the evolving pre-60S particle. Here, we show that the DEAD-box RNA helicase Spb4 with its interacting partner Rrp17 is further integrated into this maturation event. Spb4 binds to a specific class of late nucleolar pre-60S intermediates, whose cryo-EM structure revealed how its helicase activity facilitates melting and restructuring of 25S rRNA helices H62 and H63/H63a prior to Ytm1–Erb1 release. In vitro maturation of such Spb4-enriched pre-60S particles, incubated with purified Rea1 and its associated pentameric Rix1-complex in the presence of ATP, combined with cryo-EM analysis depicted the details of the Rea1-dependent large-scale pre-ribosomal remodelling. Our structural insights unveil how the Rea1 ATPase and Spb4 helicase remodel late nucleolar pre-60S particles by rRNA restructuring and dismantling of a network of several ribosomal assembly factors.

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Metadata

Authors
Mitterer, V., Thoms, M., Buschauer, R., Berninghausen, O., Hurt, E., & Beckmann, R.
Affiliations
  • Heidelberg University (DE)
  • Ludwig-Maximilians-Universität München (DE)
Corresponding author
  • Mitterer, Valentin
  • Hurt, Ed
  • Beckmann, Roland
ORCID
Venue
eLife
Publisher
openRxiv
Published
2022-11-15
Volume
12
ISSN
2050-084X (ISSN-L)
DOI
10.1101/2022.11.14.516466
PMC
PMC10154028
OpenAlex
W4327679299
Cited by
13
References
1
Citations by year
2026: 3 · 2025: 2 · 2024: 6 · 2023: 2
FWCI
1.15
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • eLife (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Deutsche Forschungsgemeinschaft (HU363/15-2)
  • European Research Council (885711-Human-Ribogenesis, ADG 741781 GLOWSOME)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nuclear Structure and Function
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
Keywords
cryo-EM structure
MeSH terms
Adenosine Triphosphatases, Ribosomal Proteins, Ribosomes, RNA Precursors, RNA, Ribosomal, Saccharomyces cerevisiae, RNA Helicases, Saccharomyces cerevisiae Proteins

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