The Npa1p complex chaperones the assembly of the earliest eukaryotic large ribosomal subunit precursor

PLoS Genetics
Clément JoretYves Henry

Abstract

The early steps of the production of the large ribosomal subunit are probably the least understood stages of eukaryotic ribosome biogenesis. The first specific precursor to the yeast large ribosomal subunit, the first pre-60S particle, contains 30 assembly factors (AFs), including 8 RNA helicases. These helicases, presumed to drive conformational rearrangements, usually lack substrate specificity in vitro. The mechanisms by which they are targeted to their correct substrate within pre-ribosomal particles and their precise molecular roles remain largely unknown. We demonstrate that the Dbp6p helicase, essential for the normal accumulation of the first pre-60S pre-ribosomal particle in S. cerevisiae, associates with a complex of four AFs, namely Npa1p, Npa2p, Nop8p and Rsa3p, prior to their incorporation into the 90S pre-ribosomal particles. By tandem affinity purifications using yeast extracts depleted of one component of the complex, we show that Npa1p forms the backbone of the complex. We provide evidence that Npa1p and Npa2p directly bind Dbp6p and we demonstrate that Npa1p is essential for the insertion of the Dbp6p helicase within 90S pre-ribosomal particles. In addition, by an in vivo cross-linking analysis (CRAC), we map ...Continue Reading

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Citations

Nov 24, 2018·Nature Reviews. Molecular Cell Biology·Sebastian Klinge, John L Woolford
Jan 24, 2019·RNA·Malik Chaker-Margot, Sebastian Klinge

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Datasets Mentioned

BETA
GSE104281

Methods Mentioned

BETA
electrophoresis
tandem affinity purifications
tandem affinity purification
size exclusion chromatography
tandem
electron microscopy
pull-down
immunoprecipitation
protein folding
two-hybrid

Software Mentioned

pyPileup
pyReadCounter

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