Implication of the box C/D snoRNP assembly factor Rsa1p in U3 snoRNP assembly

Nucleic Acids Research
Benjamin RothéBruno Charpentier

Abstract

The U3 box C/D snoRNA is one key element of 90S pre-ribosome. It contains a 5΄ domain pairing with pre-rRNA and the U3B/C and U3C΄/D motifs for U3 packaging into a unique small nucleolar ribonucleoprotein particle (snoRNP). The RNA-binding protein Snu13/SNU13 nucleates on U3B/C the assembly of box C/D proteins Nop1p/FBL and Nop56p/NOP56, and the U3-specific protein Rrp9p/U3-55K. Snu13p/SNU13 has a much lower affinity for U3C΄/D but nevertheless forms on this motif an RNP with box C/D proteins Nop1p/FBL and Nop58p/NOP58. In this study, we characterized the influence of the RNP assembly protein Rsa1 in the early steps of U3 snoRNP biogenesis in yeast and we propose a refined model of U3 snoRNP biogenesis. While recombinant Snu13p enhances the binding of Rrp9p to U3B/C, we observed that Rsa1p has no effect on this activity but forms with Snu13p and Rrp9p a U3B/C pre-RNP. In contrast, we found that Rsa1p enhances Snu13p binding on U3C΄/D. RNA footprinting experiments indicate that this positive effect most likely occurs by direct contacts of Rsa1p with the U3 snoRNA 5΄ domain. In light of the recent U3 snoRNP cryo-EM structures, our data suggest that Rsa1p has a dual role by also preventing formation of a pre-mature functional U3 RNP.

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Citations

Feb 8, 2020·Biomolecular NMR Assignments·Marie-Eve ChagotCarole Gardiennet
Jan 31, 2020·Nucleic Acids Research·Guillaume ClergetMathieu Rederstorff
Jun 3, 2021·Non-coding RNA·Laeya BaldiniStéphane Labialle
Aug 28, 2021·Current Issues in Molecular Biology·Vanessa Pinatto GasparMaria Aparecida Fernandez
Oct 24, 2017·Current Biology : CB·Ana Isabel de Los Santos-VelázquezFernando Monje-Casas

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Methods Mentioned

BETA
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electrophoresis
electrophoretic mobility shift
co-IP
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Software Mentioned

ImageQuant
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