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A proteomic screen for nucleolar SUMO targets shows SUMOylation modulates the function of Nop5/Nop58

Westman, B. J.; Verheggen, C.; Hutten, S.; Lam, Y. W.; Bertrand, E.; Lamond, A. I.

Mol Cell

2010-08-27 / vol 39 / pages 618-31

Abstract

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.

Read on PubMed

10.1016/j.molcel.2010.07.025 S1097-2765(10)00572-1 [pii]

1097-4164 (Electronic) 1097-2765 (Linking)

IGMM team(s) involved in this publication
Tags

Humans; Animals; Amino Acid Sequence; Molecular Sequence Data; Ribonucleoproteins, Small Nucleolar/genetics/*metabolism; Binding Sites; Mutation; Lysine; HeLa Cells; Rats; Recombinant Fusion Proteins/metabolism; Transfection; Nuclear Proteins/genetics/*metabolism; *Protein Processing, Post-Translational; *Proteomics/methods; Cell Nucleolus/*metabolism; Chromosomal Proteins, Non-Histone/metabolism; Ribonucleoproteins, Small Nuclear/metabolism; Small Ubiquitin-Related Modifier Proteins/*metabolism; SUMO-1 Protein/metabolism; Ubiquitins/metabolism

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