Pol5 is required for recycling of small subunit biogenesis factors and for formation of the peptide exit tunnel of the large ribosomal subunit

Nucleic Acids Res. 2020 Jan 10;48(1):405-420. doi: 10.1093/nar/gkz1079.

Abstract

More than 200 assembly factors (AFs) are required for the production of ribosomes in yeast. The stepwise association and dissociation of these AFs with the pre-ribosomal subunits occurs in a hierarchical manner to ensure correct maturation of the pre-rRNAs and assembly of the ribosomal proteins. Although decades of research have provided a wealth of insights into the functions of many AFs, others remain poorly characterized. Pol5 was initially classified with B-type DNA polymerases, however, several lines of evidence indicate the involvement of this protein in ribosome assembly. Here, we show that depletion of Pol5 affects the processing of pre-rRNAs destined for the both the large and small subunits. Furthermore, we identify binding sites for Pol5 in the 5' external transcribed spacer and within domain III of the 25S rRNA sequence. Consistent with this, we reveal that Pol5 is required for recruitment of ribosomal proteins that form the polypeptide exit tunnel in the LSU and that depletion of Pol5 impairs the release of 5' ETS fragments from early pre-40S particles. The dual functions of Pol5 in 60S assembly and recycling of pre-40S AFs suggest that this factor could contribute to ensuring the stoichiometric production of ribosomal subunits.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Binding Sites
  • DNA-Directed DNA Polymerase / chemistry
  • DNA-Directed DNA Polymerase / genetics*
  • DNA-Directed DNA Polymerase / metabolism
  • Models, Molecular
  • Nucleic Acid Conformation
  • Protein Binding
  • Protein Biosynthesis*
  • Protein Conformation, alpha-Helical
  • Protein Conformation, beta-Strand
  • Protein Interaction Domains and Motifs
  • RNA Precursors / chemistry
  • RNA Precursors / genetics
  • RNA Precursors / metabolism
  • RNA, Ribosomal / chemistry
  • RNA, Ribosomal / genetics*
  • RNA, Ribosomal / metabolism
  • Ribosome Subunits, Large, Eukaryotic / genetics*
  • Ribosome Subunits, Large, Eukaryotic / metabolism
  • Ribosome Subunits, Large, Eukaryotic / ultrastructure
  • Ribosome Subunits, Small, Eukaryotic / genetics*
  • Ribosome Subunits, Small, Eukaryotic / metabolism
  • Ribosome Subunits, Small, Eukaryotic / ultrastructure
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / genetics*
  • Saccharomyces cerevisiae Proteins / metabolism
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Thermodynamics

Substances

  • RNA Precursors
  • RNA, Ribosomal
  • Saccharomyces cerevisiae Proteins
  • RNA, ribosomal, 25S
  • DNA-Directed DNA Polymerase
  • POL5 protein, S cerevisiae