» Articles » PMID: 28959008

High-throughput RNA Structure Probing Reveals Critical Folding Events During Early 60S Ribosome Assembly in Yeast

Abstract

While the protein composition of various yeast 60S ribosomal subunit assembly intermediates has been studied in detail, little is known about ribosomal RNA (rRNA) structural rearrangements that take place during early 60S assembly steps. Using a high-throughput RNA structure probing method, we provide nucleotide resolution insights into rRNA structural rearrangements during nucleolar 60S assembly. Our results suggest that many rRNA-folding steps, such as folding of 5.8S rRNA, occur at a very specific stage of assembly, and propose that downstream nuclear assembly events can only continue once 5.8S folding has been completed. Our maps of nucleotide flexibility enable making predictions about the establishment of protein-rRNA interactions, providing intriguing insights into the temporal order of protein-rRNA as well as long-range inter-domain rRNA interactions. These data argue that many distant domains in the rRNA can assemble simultaneously during early 60S assembly and underscore the enormous complexity of 60S synthesis.Ribosome biogenesis is a dynamic process that involves the ordered assembly of ribosomal proteins and numerous RNA structural rearrangements. Here the authors apply ChemModSeq, a high-throughput RNA structure probing method, to quantitatively measure changes in RNA flexibility during the nucleolar stages of 60S assembly in yeast.

Citing Articles

The snoRNP chaperone snR190 and the Npa1 complex form a macromolecular assembly required for 60S ribosomal subunit maturation.

Hamze H, Jaafar M, Khreiss A, Dominique C, Bourdeaux J, Santo P Nucleic Acids Res. 2025; 53(5).

PMID: 40037705 PMC: 11879421. DOI: 10.1093/nar/gkaf134.


DDX RNA helicases: key players in cellular homeostasis and innate antiviral immunity.

Tapescu I, Cherry S J Virol. 2024; 98(10):e0004024.

PMID: 39212449 PMC: 11494928. DOI: 10.1128/jvi.00040-24.


Putting It All Together: The Roles of Ribosomal Proteins in Nucleolar Stages of 60S Ribosomal Assembly in the Yeast .

Ayers T, Woolford J Biomolecules. 2024; 14(8.

PMID: 39199362 PMC: 11353139. DOI: 10.3390/biom14080975.


The RNA helicase Dbp10 coordinates assembly factor association with PTC maturation during ribosome biogenesis.

Mitterer V, Hamze H, Kunowska N, Stelzl U, Henras A, Hurt E Nucleic Acids Res. 2023; 52(4):1975-1987.

PMID: 38113283 PMC: 10899779. DOI: 10.1093/nar/gkad1206.


A co-transcriptional ribosome assembly checkpoint controls nascent large ribosomal subunit maturation.

Sanghai Z, Piwowarczyk R, Vanden Broeck A, Klinge S Nat Struct Mol Biol. 2023; 30(5):594-599.

PMID: 37037974 PMC: 10191850. DOI: 10.1038/s41594-023-00947-3.


References
1.
Piir K, Tamm T, Kisly I, Tammsalu T, Remme J . Stepwise splitting of ribosomal proteins from yeast ribosomes by LiCl. PLoS One. 2014; 9(7):e101561. PMC: 4081664. DOI: 10.1371/journal.pone.0101561. View

2.
Shajani Z, Sykes M, Williamson J . Assembly of bacterial ribosomes. Annu Rev Biochem. 2011; 80:501-26. DOI: 10.1146/annurev-biochem-062608-160432. View

3.
Joseph N, Krauskopf E, Vera M, Michot B . Ribosomal internal transcribed spacer 2 (ITS2) exhibits a common core of secondary structure in vertebrates and yeast. Nucleic Acids Res. 1999; 27(23):4533-40. PMC: 148739. DOI: 10.1093/nar/27.23.4533. View

4.
Trapman J, Retel J, Planta R . Ribosomal precursor particles from yeast. Exp Cell Res. 1975; 90(1):95-104. DOI: 10.1016/0014-4827(75)90361-4. View

5.
Weis F, Giudice E, Churcher M, Jin L, Hilcenko C, Wong C . Mechanism of eIF6 release from the nascent 60S ribosomal subunit. Nat Struct Mol Biol. 2015; 22(11):914-9. PMC: 4871238. DOI: 10.1038/nsmb.3112. View