A proteomic screen for nucleolar SUMO targets shows SUMOylation modulates the function of Nop5/Nop58
| Autor: | Yun Wah Lam, Belinda J. Westman, Angus I. Lamond, Edouard Bertrand, Céline Verheggen, Saskia Hutten |
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| Přispěvatelé: | Institut de Génétique Moléculaire de Montpellier (IGMM), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM) |
| Rok vydání: | 2010 |
| Předmět: |
Proteomics
Nucleolus Chromosomal Proteins Non-Histone PROTEINS Recombinant Fusion Proteins Quantitative proteomics Molecular Sequence Data SUMO-1 Protein SUMO protein Post-Translational *Proteomics/methods Rats Recombinant Fusion Proteins/metabolism Ribonucleoproteins Biology Transfection Article 03 medical and health sciences Ribonucleoproteins Small Nucleolar Stable isotope labeling by amino acids in cell culture medicine Animals Humans [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Amino Acid Sequence Small nucleolar RNA Nuclear protein Molecular Biology Ubiquitins 030304 developmental biology Small Nucleolar/genetics/*metabolism SUMO-1 Protein/metabolism Small Ubiquitin-Related Modifier Proteins/*metabolism Transfection Ubiquitins/metabolism Genetics Small Nuclear/metabolism Ribonucleoproteins 0303 health sciences Binding Sites Lysine 030302 biochemistry & molecular biology Nuclear Proteins Cell Biology Amino Acid Sequence Animals Binding Sites Cell Nucleolus/*metabolism Chromosomal Proteins Ribonucleoproteins Small Nuclear Cell biology Rats Cell nucleus medicine.anatomical_structure Ribosome Subunits Mutation Small Ubiquitin-Related Modifier Proteins RNA Protein Processing Post-Translational Cell Nucleolus Non-Histone/metabolism HeLa Cells Humans Lysine Molecular Sequence Data Mutation Nuclear Proteins/genetics/*metabolism *Protein Processing HeLa Cells |
| Zdroj: | Molecular Cell Molecular Cell; Vol 39 Molecular Cell, Elsevier, 2010, 39 (4), pp.618--31. ⟨10.1016/j.molcel.2010.07.025⟩ |
| ISSN: | 1097-4164 1097-2765 |
| DOI: | 10.1016/j.molcel.2010.07.025⟩ |
| Popis: | Summary Posttranslational SUMO modification is an important mechanism of regulating protein function, especially in the cell nucleus. The nucleolus is the subnuclear organelle responsible for rRNA synthesis, processing, and assembly of the large and small ribosome subunits. Here, we have used SILAC-based quantitative proteomics to identify nucleolar SUMOylated proteins. This reveals a role for SUMOylation in the biogenesis and/or function of small nucleolar ribonucleoprotein complexes (snoRNPs) via the targeting of Nhp2 and Nop58. Using combined in vitro and in vivo approaches, both Nhp2 and Nop58 (also known as Nop5) are shown to be substrates for SUMOylation. Mutational analyses revealed the sites of modification on Nhp2 as K5, and on Nop58 as K467 and K497. Unlike Nop58 and Nhp2, the closely related Nop56 and 15.5K proteins appear not to be SUMO targets. SUMOylation is essential for high-affinity Nop58 binding to snoRNAs. This study provides direct evidence linking SUMO modification with snoRNP function. Highlights ► Nucleolar SUMO targets identified using SILAC-based quantitative proteomics ► K5 in Nhp2 and K467/K497 in Nop58 are SUMOylated both in vitro and in vivo ► SUMOylation may explain the distinct roles of Nop56/Nop58 and Nhp2/15.5K in snoRNPs ► SUMOylation is important for high-affinity binding of box C/D snoRNPs to snoRNAs |
| Databáze: | OpenAIRE |
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