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Arginine methylation of the C-terminus RGG motif promotes TOP3B topoisomerase activity and stress granule localization
DNA topoisomerase 3B (TOP3B) is unique among all mammalian topoisomerases for its dual activities that resolve both DNA and RNA topological entanglements to facilitate transcription and translation. However, the mechanism by which TOP3B activity is regulated is still elusive. Here, we have identifie...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Oxford University Press
2018
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5888246/ https://www.ncbi.nlm.nih.gov/pubmed/29471495 http://dx.doi.org/10.1093/nar/gky103 |
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author | Huang, Lifeng Wang, Zhihao Narayanan, Nithya Yang, Yanzhong |
author_facet | Huang, Lifeng Wang, Zhihao Narayanan, Nithya Yang, Yanzhong |
author_sort | Huang, Lifeng |
collection | PubMed |
description | DNA topoisomerase 3B (TOP3B) is unique among all mammalian topoisomerases for its dual activities that resolve both DNA and RNA topological entanglements to facilitate transcription and translation. However, the mechanism by which TOP3B activity is regulated is still elusive. Here, we have identified arginine methylation as an important post-translational modification (PTM) for TOP3B activity. Protein arginine methyltransferase (PRMT) 1, PRMT3 and PRMT6 all methylate TOP3B in vitro at its C-terminal arginine (R) and glycine (G)-rich motif. Site-directed mutagenesis analysis identified R833 and R835 as the major methylation sites. Using a methylation-specific antibody, we confirmed that TOP3B is methylated in cells and that mutation of R833 and R835 to lysine (K) significantly reduces TOP3B methylation. The methylation-deficient TOP3B (R833/835K) is less active in resolving negatively supercoiled DNA, which consequently lead to accumulation of co-transcriptionally formed R-loops in vitro and in cells. Additionally, the methylation-deficient TOP3B (R833/835K) shows reduced stress granule localization, indicating that methylation is critical for TOP3B function in translation regulation. Mechanistically, we found that R833/835 methylation is partially involved in the interaction of TOP3B with its auxiliary factor, the Tudor domain-containing protein 3 (TDRD3). Together, our findings provide the first evidence for the regulation of TOP3B activity by PTM. |
format | Online Article Text |
id | pubmed-5888246 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-58882462018-04-11 Arginine methylation of the C-terminus RGG motif promotes TOP3B topoisomerase activity and stress granule localization Huang, Lifeng Wang, Zhihao Narayanan, Nithya Yang, Yanzhong Nucleic Acids Res Molecular Biology DNA topoisomerase 3B (TOP3B) is unique among all mammalian topoisomerases for its dual activities that resolve both DNA and RNA topological entanglements to facilitate transcription and translation. However, the mechanism by which TOP3B activity is regulated is still elusive. Here, we have identified arginine methylation as an important post-translational modification (PTM) for TOP3B activity. Protein arginine methyltransferase (PRMT) 1, PRMT3 and PRMT6 all methylate TOP3B in vitro at its C-terminal arginine (R) and glycine (G)-rich motif. Site-directed mutagenesis analysis identified R833 and R835 as the major methylation sites. Using a methylation-specific antibody, we confirmed that TOP3B is methylated in cells and that mutation of R833 and R835 to lysine (K) significantly reduces TOP3B methylation. The methylation-deficient TOP3B (R833/835K) is less active in resolving negatively supercoiled DNA, which consequently lead to accumulation of co-transcriptionally formed R-loops in vitro and in cells. Additionally, the methylation-deficient TOP3B (R833/835K) shows reduced stress granule localization, indicating that methylation is critical for TOP3B function in translation regulation. Mechanistically, we found that R833/835 methylation is partially involved in the interaction of TOP3B with its auxiliary factor, the Tudor domain-containing protein 3 (TDRD3). Together, our findings provide the first evidence for the regulation of TOP3B activity by PTM. Oxford University Press 2018-04-06 2018-02-19 /pmc/articles/PMC5888246/ /pubmed/29471495 http://dx.doi.org/10.1093/nar/gky103 Text en © The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Molecular Biology Huang, Lifeng Wang, Zhihao Narayanan, Nithya Yang, Yanzhong Arginine methylation of the C-terminus RGG motif promotes TOP3B topoisomerase activity and stress granule localization |
title | Arginine methylation of the C-terminus RGG motif promotes TOP3B topoisomerase activity and stress granule localization |
title_full | Arginine methylation of the C-terminus RGG motif promotes TOP3B topoisomerase activity and stress granule localization |
title_fullStr | Arginine methylation of the C-terminus RGG motif promotes TOP3B topoisomerase activity and stress granule localization |
title_full_unstemmed | Arginine methylation of the C-terminus RGG motif promotes TOP3B topoisomerase activity and stress granule localization |
title_short | Arginine methylation of the C-terminus RGG motif promotes TOP3B topoisomerase activity and stress granule localization |
title_sort | arginine methylation of the c-terminus rgg motif promotes top3b topoisomerase activity and stress granule localization |
topic | Molecular Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5888246/ https://www.ncbi.nlm.nih.gov/pubmed/29471495 http://dx.doi.org/10.1093/nar/gky103 |
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