Two orthogonal cleavages separate subunit RNAs in mouse ribosome biogenesis

Minshi Wang, Leonid Anikin, Dimitri G. Pestov

Research output: Contribution to journalArticlepeer-review

47 Scopus citations

Abstract

Ribosome biogenesis is a dynamic multistep process, many features of which are still incompletely documented. Here, we show that changes in this pathway can be captured and annotated by means of a graphic set of pre-rRNA ratios, a technique we call Ratio Analysis of Multiple Precursors (RAMP). We find that knocking down a ribosome synthesis factor produces a characteristic RAMP profile that exhibits consistency across a range of depletion levels. This facilitates the inference of affected steps and simplifies comparative analysis. We applied RAMP to examine how endonucleolytic cleavages of the mouse pre-rRNA transcript in the internal transcribed spacer 1 (ITS1) are affected by depletion of factors required for maturation of the small ribosomal sub-unit (Rcl1, Fcf1/Utp24, Utp23) and the large sub-unit (Pes1, Nog1). The data suggest that completion of early maturation in a subunit triggers its release from the common pre-rRNA transcript by stimulating cleavage at the proximal site in ITS1. We also find that splitting of pre-rRNA in the 3′ region of ITS1 is prevalent in adult mouse tissues and quiescent cells, as it is in human cells. We propose a model for sub-unit separation during mammalian ribosome synthesis and discuss its implications for understanding pre-rRNA processing pathways.

Original languageEnglish (US)
Pages (from-to)11180-11191
Number of pages12
JournalNucleic acids research
Volume42
Issue number17
DOIs
StatePublished - Jun 27 2014

All Science Journal Classification (ASJC) codes

  • Genetics

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