Force measurements show that uL4 and uL24 mechanically stabilize a fragment of 23S rRNA essential for ribosome assembly

Laurent Geffroy, Thierry Bizebard, Ryo Aoyama, Takuya Ueda, Ulrich Bockelmann

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

In vitro reconstitution studies have shown that ribosome assembly is highly cooperative and starts with the binding of a few ribosomal (r-) proteins to rRNA. It is unknown how these early binders act. Focusing on the initial stage of the assembly of the large subunit of the Escherichia coli ribosome, we prepared a 79-nucleotide-long region of 23S rRNA encompassing the binding sites of the early binders uL4 and uL24. Force signals were measured in a DNA/RNA dumbbell configuration with a double optical tweezers setup. The rRNA fragment was stretched until unfolded, in the absence or in the presence of the r-proteins (either uL4, uL24, or both). We show that the r-proteins uL4 and uL24 individually stabilize the rRNA fragment, both acting as molecular clamps. Interestingly, this mechanical stabilization is enhanced when both proteins are bound simultaneously. Independently, we observe a cooperative binding of uL4 and uL24 to the rRNA fragment. These two aspects of r-proteins binding both contribute to the efficient stabilization of the 3D structure of the rRNA fragment under investigation. We finally consider implications of our results for large ribosomal subunit assembly.

Original languageEnglish
Pages (from-to)472-480
Number of pages9
JournalRNA
Volume25
Issue number4
DOIs
Publication statusPublished - 2019 Apr 1

Keywords

  • Optical trap
  • Ribosome assembly
  • Single molecule

ASJC Scopus subject areas

  • Molecular Biology

Fingerprint Dive into the research topics of 'Force measurements show that uL4 and uL24 mechanically stabilize a fragment of 23S rRNA essential for ribosome assembly'. Together they form a unique fingerprint.

  • Cite this