Proteomic and 3D structure analyses highlight the C/D box snoRNP assembly mechanism and its control

Jonathan Bizarro, Christophe Charron, Séverine Boulon, Belinda Westman, Bérengère Pradet-Balade, Franck Vandermoere, Marie Eve Chagot, Marie Hallais, Yasmeen Ahmad, Heinrich Leonhardt, Angus Lamond, Xavier Manival, Christiane Branlant, Bruno Charpentier, Céline Verheggen, Edouard Bertrand

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

In vitro, assembly of box C/D small nucleolar ribonucleoproteins (snoRNPs) involves the sequential recruitment of core proteins to snoRNAs. In vivo, however, assembly factors are required (NUFIP, BCD1, and the HSP90-R2TP complex), and it is unknown whether a similar sequential scheme applies. In this paper, we describe systematic quantitative stable isotope labeling by amino acids in cell culture proteomic experiments and the crystal structure of the core protein Snu13p/15.5K bound to a fragment of the assembly factor Rsa1p/NUFIP. This revealed several unexpected features: (a) the existence of a protein-only pre-snoRNP complex containing five assembly factors and two core proteins, 15.5K and Nop58; (b) the characterization of ZNHIT3, which is present in the protein-only complex but gets released upon binding to C/D snoRNAs; (c) the dynamics of the R2TP complex, which appears to load/unload RuvBL AAA+ adenosine triphosphatase from pre-snoRNPs; and (d) a potential mechanism for preventing premature activation of snoRNP catalytic activity. These data provide a framework for understanding the assembly of box C/D snoRNPs.

Original languageEnglish (US)
Pages (from-to)463-480
Number of pages18
JournalJournal of Cell Biology
Volume207
Issue number4
DOIs
StatePublished - Jan 1 2014
Externally publishedYes

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Small Nucleolar Ribonucleoproteins
Proteomics
Small Nucleolar RNA
Proteins
Isotope Labeling
Adenosine Triphosphatases
Cell Culture Techniques
Amino Acids

ASJC Scopus subject areas

  • Cell Biology

Cite this

Bizarro, J., Charron, C., Boulon, S., Westman, B., Pradet-Balade, B., Vandermoere, F., ... Bertrand, E. (2014). Proteomic and 3D structure analyses highlight the C/D box snoRNP assembly mechanism and its control. Journal of Cell Biology, 207(4), 463-480. https://doi.org/10.1083/jcb.201404160

Proteomic and 3D structure analyses highlight the C/D box snoRNP assembly mechanism and its control. / Bizarro, Jonathan; Charron, Christophe; Boulon, Séverine; Westman, Belinda; Pradet-Balade, Bérengère; Vandermoere, Franck; Chagot, Marie Eve; Hallais, Marie; Ahmad, Yasmeen; Leonhardt, Heinrich; Lamond, Angus; Manival, Xavier; Branlant, Christiane; Charpentier, Bruno; Verheggen, Céline; Bertrand, Edouard.

In: Journal of Cell Biology, Vol. 207, No. 4, 01.01.2014, p. 463-480.

Research output: Contribution to journalArticle

Bizarro, J, Charron, C, Boulon, S, Westman, B, Pradet-Balade, B, Vandermoere, F, Chagot, ME, Hallais, M, Ahmad, Y, Leonhardt, H, Lamond, A, Manival, X, Branlant, C, Charpentier, B, Verheggen, C & Bertrand, E 2014, 'Proteomic and 3D structure analyses highlight the C/D box snoRNP assembly mechanism and its control', Journal of Cell Biology, vol. 207, no. 4, pp. 463-480. https://doi.org/10.1083/jcb.201404160
Bizarro, Jonathan ; Charron, Christophe ; Boulon, Séverine ; Westman, Belinda ; Pradet-Balade, Bérengère ; Vandermoere, Franck ; Chagot, Marie Eve ; Hallais, Marie ; Ahmad, Yasmeen ; Leonhardt, Heinrich ; Lamond, Angus ; Manival, Xavier ; Branlant, Christiane ; Charpentier, Bruno ; Verheggen, Céline ; Bertrand, Edouard. / Proteomic and 3D structure analyses highlight the C/D box snoRNP assembly mechanism and its control. In: Journal of Cell Biology. 2014 ; Vol. 207, No. 4. pp. 463-480.
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