Puf6 primes 60S pre-ribosome nuclear export at low temperature

• Published in: Nature communications Vol. 12; no. 1; p. 4696
• Main Authors: Gerhardy, Stefan, Oborská-Oplová, Michaela, Gillet, Ludovic, Börner, Richard, van Nues, Rob, Leitner, Alexander, Michel, Erich, Petkowski, Janusz J, Granneman, Sander, Sigel, Roland K O, Aebersold, Ruedi, Panse, Vikram Govind
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 04.08.2021; Nature Publishing Group UK; Nature Portfolio
• Subjects: Active Transport, Cell Nucleus; Assembly; Cell Nucleus - metabolism; Cold Temperature; Compaction; Critical temperature; Energy transfer; Folding; GTP Phosphohydrolases - metabolism; Intermediates; Low temperature; Magnesium; Mutation; Nuclear transport; Protein Binding; Protein Interaction Domains and Motifs; Protein structure; Proteome - metabolism; Proteomes; Ribosome Subunits, Large, Eukaryotic - chemistry; Ribosome Subunits, Large, Eukaryotic - metabolism; Ribosomes; Ribosomes - metabolism; RNA Folding; RNA Precursors - chemistry; RNA Precursors - metabolism; RNA, Ribosomal - chemistry; RNA, Ribosomal - metabolism; RNA-Binding Proteins - chemistry; RNA-Binding Proteins - genetics; RNA-Binding Proteins - metabolism; rRNA; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae - physiology; Saccharomyces cerevisiae Proteins - chemistry; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; Secondary structure; Yeast; Yeasts
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-24964-2

Productive ribosomal RNA (rRNA) compaction during ribosome assembly necessitates establishing correct tertiary contacts between distant secondary structure elements. Here, we quantify the response of the yeast proteome to low temperature (LT), a condition where aberrant mis-paired RNA folding intermediates accumulate. We show that, at LT, yeast cells globally boost production of their ribosome assembly machinery. We find that the LT-induced assembly factor, Puf6, binds to the nascent catalytic RNA-rich subunit interface within the 60S pre-ribosome, at a site that eventually loads the nuclear export apparatus. Ensemble Förster resonance energy transfer studies show that Puf6 mimics the role of Mg to usher a unique long-range tertiary contact to compact rRNA. At LT, puf6 mutants accumulate 60S pre-ribosomes in the nucleus, thus unveiling Puf6-mediated rRNA compaction as a critical temperature-regulated rescue mechanism that counters rRNA misfolding to prime export competence.