TY - JOUR
T1 - Proteomic and 3D structure analyses highlight the C/D box snoRNP assembly mechanism and its control
AU - Bizarro, Jonathan
AU - Charron, Christophe
AU - Boulon, Séverine
AU - Westman, Belinda
AU - Pradet-Balade, Bérengère
AU - Vandermoere, Franck
AU - Chagot, Marie Eve
AU - Hallais, Marie
AU - Ahmad, Yasmeen
AU - Leonhardt, Heinrich
AU - Lamond, Angus
AU - Manival, Xavier
AU - Branlant, Christiane
AU - Charpentier, Bruno
AU - Verheggen, Céline
AU - Bertrand, Edouard
N1 - Publisher Copyright:
© 2014 Bizarro et al.
PY - 2014
Y1 - 2014
N2 - 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.
AB - 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.
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U2 - 10.1083/jcb.201404160
DO - 10.1083/jcb.201404160
M3 - Article
C2 - 25404746
AN - SCOPUS:84918533144
SN - 0021-9525
VL - 207
SP - 463
EP - 480
JO - Journal of Cell Biology
JF - Journal of Cell Biology
IS - 4
ER -