A Proteomic Screen for Nucleolar SUMO Targets Shows SUMOylation Modulates the Function of Nop5/Nop58
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 qu...
Autores principales: | , , , , , |
---|---|
Formato: | Texto |
Lenguaje: | English |
Publicado: |
Cell Press
2010
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2938476/ https://www.ncbi.nlm.nih.gov/pubmed/20797632 http://dx.doi.org/10.1016/j.molcel.2010.07.025 |
_version_ | 1782186625588002816 |
---|---|
author | Westman, Belinda J. Verheggen, Céline Hutten, Saskia Lam, Yun Wah Bertrand, Edouard Lamond, Angus I. |
author_facet | Westman, Belinda J. Verheggen, Céline Hutten, Saskia Lam, Yun Wah Bertrand, Edouard Lamond, Angus I. |
author_sort | Westman, Belinda J. |
collection | PubMed |
description | 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. |
format | Text |
id | pubmed-2938476 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Cell Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-29384762010-10-13 A Proteomic Screen for Nucleolar SUMO Targets Shows SUMOylation Modulates the Function of Nop5/Nop58 Westman, Belinda J. Verheggen, Céline Hutten, Saskia Lam, Yun Wah Bertrand, Edouard Lamond, Angus I. Mol Cell Article 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. Cell Press 2010-08-27 /pmc/articles/PMC2938476/ /pubmed/20797632 http://dx.doi.org/10.1016/j.molcel.2010.07.025 Text en © 2010 ELL & Excerpta Medica. https://creativecommons.org/licenses/by/3.0/ Open Access under CC BY 3.0 (https://creativecommons.org/licenses/by/3.0/) license |
spellingShingle | Article Westman, Belinda J. Verheggen, Céline Hutten, Saskia Lam, Yun Wah Bertrand, Edouard Lamond, Angus I. A Proteomic Screen for Nucleolar SUMO Targets Shows SUMOylation Modulates the Function of Nop5/Nop58 |
title | A Proteomic Screen for Nucleolar SUMO Targets Shows SUMOylation Modulates the Function of Nop5/Nop58 |
title_full | A Proteomic Screen for Nucleolar SUMO Targets Shows SUMOylation Modulates the Function of Nop5/Nop58 |
title_fullStr | A Proteomic Screen for Nucleolar SUMO Targets Shows SUMOylation Modulates the Function of Nop5/Nop58 |
title_full_unstemmed | A Proteomic Screen for Nucleolar SUMO Targets Shows SUMOylation Modulates the Function of Nop5/Nop58 |
title_short | A Proteomic Screen for Nucleolar SUMO Targets Shows SUMOylation Modulates the Function of Nop5/Nop58 |
title_sort | proteomic screen for nucleolar sumo targets shows sumoylation modulates the function of nop5/nop58 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2938476/ https://www.ncbi.nlm.nih.gov/pubmed/20797632 http://dx.doi.org/10.1016/j.molcel.2010.07.025 |
work_keys_str_mv | AT westmanbelindaj aproteomicscreenfornucleolarsumotargetsshowssumoylationmodulatesthefunctionofnop5nop58 AT verheggenceline aproteomicscreenfornucleolarsumotargetsshowssumoylationmodulatesthefunctionofnop5nop58 AT huttensaskia aproteomicscreenfornucleolarsumotargetsshowssumoylationmodulatesthefunctionofnop5nop58 AT lamyunwah aproteomicscreenfornucleolarsumotargetsshowssumoylationmodulatesthefunctionofnop5nop58 AT bertrandedouard aproteomicscreenfornucleolarsumotargetsshowssumoylationmodulatesthefunctionofnop5nop58 AT lamondangusi aproteomicscreenfornucleolarsumotargetsshowssumoylationmodulatesthefunctionofnop5nop58 AT westmanbelindaj proteomicscreenfornucleolarsumotargetsshowssumoylationmodulatesthefunctionofnop5nop58 AT verheggenceline proteomicscreenfornucleolarsumotargetsshowssumoylationmodulatesthefunctionofnop5nop58 AT huttensaskia proteomicscreenfornucleolarsumotargetsshowssumoylationmodulatesthefunctionofnop5nop58 AT lamyunwah proteomicscreenfornucleolarsumotargetsshowssumoylationmodulatesthefunctionofnop5nop58 AT bertrandedouard proteomicscreenfornucleolarsumotargetsshowssumoylationmodulatesthefunctionofnop5nop58 AT lamondangusi proteomicscreenfornucleolarsumotargetsshowssumoylationmodulatesthefunctionofnop5nop58 |