A role for SUMOylation in snoRNP biogenesis revealed by quantitative proteomics

Belinda J. Westman, Angus I. Lamond

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

A role for SUMOylation in the biogenesis and/or function of Box C/D snoRNPs has been reported, mediated via SUMO2 conjugation to the core snoRNP protein, Nop58. A quantitative proteomics screen, based on SILAC (stable-isotope labeling by amino acids in cell culture) and mass spectrometry using extracts prepared from cultured mammalian cells expressing either 6His-SUMO1 or -SUMO2, revealed that the snoRNP-related proteins Nop58, Nhp2, DKC1 and NOLC1 are amongst the main SUMO-modified proteins in the nucleolus. SUMOylation of Nhp2 and endogenous Nop58 was confirmed using a combination of in vitro and cell-based assays and the modified lysines identified by site-directed mutagenesis. SUMOylation of Nop58 was found to be important for high-affinity Box C/D snoRNA binding and the localization of newly transcribed snoRNAs to the nucleolus. Here, these findings are reviewed and a model for understanding Nop58 SUMOylation in the context of Box C/D snoRNP biogenesis is presented. Given the essential role of snoRNPs in the modification of pre-ribosomal RNA, this work suggests that SUMO, snoRNPs and ribosome assembly, and thus cellular translation, growth and proliferation, may be linked via novel mechanisms which warrant further investigation.
Original languageEnglish
Pages (from-to)30-37
Number of pages8
JournalNucleus
Volume2
Issue number1
DOIs
Publication statusPublished - 1 Jan 2011

Fingerprint

Small Nucleolar Ribonucleoproteins
Sumoylation
Proteomics
Small Nucleolar RNA
Small Ubiquitin-Related Modifier Proteins
Isotope Labeling
Ribosomal RNA
Site-Directed Mutagenesis
Ribosomes
Lysine
Cultured Cells
Mass Spectrometry
Proteins
Cell Culture Techniques
Amino Acids
Growth

Cite this

@article{d70b029d66944e6b822813d9e0a6388d,
title = "A role for SUMOylation in snoRNP biogenesis revealed by quantitative proteomics",
abstract = "A role for SUMOylation in the biogenesis and/or function of Box C/D snoRNPs has been reported, mediated via SUMO2 conjugation to the core snoRNP protein, Nop58. A quantitative proteomics screen, based on SILAC (stable-isotope labeling by amino acids in cell culture) and mass spectrometry using extracts prepared from cultured mammalian cells expressing either 6His-SUMO1 or -SUMO2, revealed that the snoRNP-related proteins Nop58, Nhp2, DKC1 and NOLC1 are amongst the main SUMO-modified proteins in the nucleolus. SUMOylation of Nhp2 and endogenous Nop58 was confirmed using a combination of in vitro and cell-based assays and the modified lysines identified by site-directed mutagenesis. SUMOylation of Nop58 was found to be important for high-affinity Box C/D snoRNA binding and the localization of newly transcribed snoRNAs to the nucleolus. Here, these findings are reviewed and a model for understanding Nop58 SUMOylation in the context of Box C/D snoRNP biogenesis is presented. Given the essential role of snoRNPs in the modification of pre-ribosomal RNA, this work suggests that SUMO, snoRNPs and ribosome assembly, and thus cellular translation, growth and proliferation, may be linked via novel mechanisms which warrant further investigation.",
author = "Westman, {Belinda J.} and Lamond, {Angus I.}",
note = "MEDLINE{\circledR} is the source for the MeSH terms of this document.",
year = "2011",
month = "1",
day = "1",
doi = "10.4161/nucl.2.1.14437",
language = "English",
volume = "2",
pages = "30--37",
journal = "Nucleus",
issn = "1949-1034",
publisher = "Taylor & Francis",
number = "1",

}

A role for SUMOylation in snoRNP biogenesis revealed by quantitative proteomics. / Westman, Belinda J.; Lamond, Angus I.

In: Nucleus, Vol. 2, No. 1, 01.01.2011, p. 30-37.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A role for SUMOylation in snoRNP biogenesis revealed by quantitative proteomics

AU - Westman, Belinda J.

AU - Lamond, Angus I.

N1 - MEDLINE® is the source for the MeSH terms of this document.

PY - 2011/1/1

Y1 - 2011/1/1

N2 - A role for SUMOylation in the biogenesis and/or function of Box C/D snoRNPs has been reported, mediated via SUMO2 conjugation to the core snoRNP protein, Nop58. A quantitative proteomics screen, based on SILAC (stable-isotope labeling by amino acids in cell culture) and mass spectrometry using extracts prepared from cultured mammalian cells expressing either 6His-SUMO1 or -SUMO2, revealed that the snoRNP-related proteins Nop58, Nhp2, DKC1 and NOLC1 are amongst the main SUMO-modified proteins in the nucleolus. SUMOylation of Nhp2 and endogenous Nop58 was confirmed using a combination of in vitro and cell-based assays and the modified lysines identified by site-directed mutagenesis. SUMOylation of Nop58 was found to be important for high-affinity Box C/D snoRNA binding and the localization of newly transcribed snoRNAs to the nucleolus. Here, these findings are reviewed and a model for understanding Nop58 SUMOylation in the context of Box C/D snoRNP biogenesis is presented. Given the essential role of snoRNPs in the modification of pre-ribosomal RNA, this work suggests that SUMO, snoRNPs and ribosome assembly, and thus cellular translation, growth and proliferation, may be linked via novel mechanisms which warrant further investigation.

AB - A role for SUMOylation in the biogenesis and/or function of Box C/D snoRNPs has been reported, mediated via SUMO2 conjugation to the core snoRNP protein, Nop58. A quantitative proteomics screen, based on SILAC (stable-isotope labeling by amino acids in cell culture) and mass spectrometry using extracts prepared from cultured mammalian cells expressing either 6His-SUMO1 or -SUMO2, revealed that the snoRNP-related proteins Nop58, Nhp2, DKC1 and NOLC1 are amongst the main SUMO-modified proteins in the nucleolus. SUMOylation of Nhp2 and endogenous Nop58 was confirmed using a combination of in vitro and cell-based assays and the modified lysines identified by site-directed mutagenesis. SUMOylation of Nop58 was found to be important for high-affinity Box C/D snoRNA binding and the localization of newly transcribed snoRNAs to the nucleolus. Here, these findings are reviewed and a model for understanding Nop58 SUMOylation in the context of Box C/D snoRNP biogenesis is presented. Given the essential role of snoRNPs in the modification of pre-ribosomal RNA, this work suggests that SUMO, snoRNPs and ribosome assembly, and thus cellular translation, growth and proliferation, may be linked via novel mechanisms which warrant further investigation.

UR - http://www.scopus.com/inward/record.url?scp=79952679740&partnerID=8YFLogxK

U2 - 10.4161/nucl.2.1.14437

DO - 10.4161/nucl.2.1.14437

M3 - Article

VL - 2

SP - 30

EP - 37

JO - Nucleus

JF - Nucleus

SN - 1949-1034

IS - 1

ER -