Stepwise and dynamic assembly of the earliest precursors of small ribosomal subunits in yeast

Abstract

In this study, Zhang et al. researched how the 90S preribosomal particle is cotranscriptionally assembled in yeast using a novel approach. They determined the assembly point of 65 proteins and the U3, U14, and snR30 snoRNAs, revealing a stepwise and dynamic assembly map, thereby advancing our understanding of small subunit biogenesis.

Abstract

The eukaryotic ribosomal RNA (rRNA) is associated cotranscriptionally with numerous factors into an enormous 90S preribosomal particle that conducts early processing of small ribosomal subunits. The assembly pathway and structure of the 90S particle is poorly understood. Here, we affinity-purified and analyzed the constituents of yeast 90S particles that were assembled on a series of plasmid-encoded 3′-truncated pre-18S RNAs. We determined the assembly point of 65 proteins and the U3, U14, and snR30 small nucleolar RNAs (snoRNAs), revealing a stepwise and dynamic assembly map. The 5′ external transcribed spacer (ETS) alone can nucleate a large complex. When the 18S rRNA is nearly complete, the 90S structure undergoes a dramatic reorganization, releasing U14, snR30, and 14 protein factors that bind earlier. We also identified a reference state of 90S that is fully assembled yet has not undergone 5′ETS processing. The assembly map present here provides a new framework to understand small subunit biogenesis.