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SUMO Chain Formation Is Required for Response to Replication Arrest in S. pombe

SUMO is a ubiquitin-like protein that is post-translationally attached to one or more lysine residues on target proteins. Despite having only 18% sequence identity with ubiquitin, SUMO contains the conserved ββαββαβ fold present in ubiquitin. However, SUMO differs from ubiquitin in having an extende...

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Detalles Bibliográficos
Autores principales: Skilton, Andrew, Ho, Jenny C. Y., Mercer, Brenda, Outwin, Emily, Watts, Felicity Z.
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2727700/
https://www.ncbi.nlm.nih.gov/pubmed/19707600
http://dx.doi.org/10.1371/journal.pone.0006750
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author Skilton, Andrew
Ho, Jenny C. Y.
Mercer, Brenda
Outwin, Emily
Watts, Felicity Z.
author_facet Skilton, Andrew
Ho, Jenny C. Y.
Mercer, Brenda
Outwin, Emily
Watts, Felicity Z.
author_sort Skilton, Andrew
collection PubMed
description SUMO is a ubiquitin-like protein that is post-translationally attached to one or more lysine residues on target proteins. Despite having only 18% sequence identity with ubiquitin, SUMO contains the conserved ββαββαβ fold present in ubiquitin. However, SUMO differs from ubiquitin in having an extended N-terminus. In S. pombe the N-terminus of SUMO/Pmt3 is significantly longer than those of SUMO in S. cerevisiae, human and Drosophila. Here we investigate the role of this N-terminal region. We have used two dimensional gel electrophoresis to demonstrate that S. pombe SUMO/Pmt3 is phosphorylated, and that this occurs on serine residues at the extreme N-terminus of the protein. Mutation of these residues (in pmt3-1) results in a dramatic reduction in both the levels of high Mr SUMO-containing species and of total SUMO/Pmt3, indicating that phosphorylation of SUMO/Pmt3 is required for its stability. Despite the significant reduction in high Mr SUMO-containing species, pmt3-1 cells do not display an aberrant cell morphology or sensitivity to genotoxins or stress. Additionally, we demonstrate that two lysine residues in the N-terminus of S. pombe SUMO/Pmt3 (K14 and K30) can act as acceptor sites for SUMO chain formation in vitro. Inability to form SUMO chains results in aberrant cell and nuclear morphologies, including stretched and fragmented chromatin. SUMO chain mutants are sensitive to the DNA synthesis inhibitor, hydroxyurea (HU), but not to other genotoxins, such as UV, MMS or CPT. This implies a role for SUMO chains in the response to replication arrest in S. pombe.
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spelling pubmed-27277002009-08-25 SUMO Chain Formation Is Required for Response to Replication Arrest in S. pombe Skilton, Andrew Ho, Jenny C. Y. Mercer, Brenda Outwin, Emily Watts, Felicity Z. PLoS One Research Article SUMO is a ubiquitin-like protein that is post-translationally attached to one or more lysine residues on target proteins. Despite having only 18% sequence identity with ubiquitin, SUMO contains the conserved ββαββαβ fold present in ubiquitin. However, SUMO differs from ubiquitin in having an extended N-terminus. In S. pombe the N-terminus of SUMO/Pmt3 is significantly longer than those of SUMO in S. cerevisiae, human and Drosophila. Here we investigate the role of this N-terminal region. We have used two dimensional gel electrophoresis to demonstrate that S. pombe SUMO/Pmt3 is phosphorylated, and that this occurs on serine residues at the extreme N-terminus of the protein. Mutation of these residues (in pmt3-1) results in a dramatic reduction in both the levels of high Mr SUMO-containing species and of total SUMO/Pmt3, indicating that phosphorylation of SUMO/Pmt3 is required for its stability. Despite the significant reduction in high Mr SUMO-containing species, pmt3-1 cells do not display an aberrant cell morphology or sensitivity to genotoxins or stress. Additionally, we demonstrate that two lysine residues in the N-terminus of S. pombe SUMO/Pmt3 (K14 and K30) can act as acceptor sites for SUMO chain formation in vitro. Inability to form SUMO chains results in aberrant cell and nuclear morphologies, including stretched and fragmented chromatin. SUMO chain mutants are sensitive to the DNA synthesis inhibitor, hydroxyurea (HU), but not to other genotoxins, such as UV, MMS or CPT. This implies a role for SUMO chains in the response to replication arrest in S. pombe. Public Library of Science 2009-08-25 /pmc/articles/PMC2727700/ /pubmed/19707600 http://dx.doi.org/10.1371/journal.pone.0006750 Text en Skilton et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Skilton, Andrew
Ho, Jenny C. Y.
Mercer, Brenda
Outwin, Emily
Watts, Felicity Z.
SUMO Chain Formation Is Required for Response to Replication Arrest in S. pombe
title SUMO Chain Formation Is Required for Response to Replication Arrest in S. pombe
title_full SUMO Chain Formation Is Required for Response to Replication Arrest in S. pombe
title_fullStr SUMO Chain Formation Is Required for Response to Replication Arrest in S. pombe
title_full_unstemmed SUMO Chain Formation Is Required for Response to Replication Arrest in S. pombe
title_short SUMO Chain Formation Is Required for Response to Replication Arrest in S. pombe
title_sort sumo chain formation is required for response to replication arrest in s. pombe
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2727700/
https://www.ncbi.nlm.nih.gov/pubmed/19707600
http://dx.doi.org/10.1371/journal.pone.0006750
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