Detection of RNA-DNA association by a proximity ligation-based method

We describe a proximity ligation assay (PLA)-based method of assessing association of DNA and RNA in single cells during the cell cycle. Pulse-labeling of DNA with EdU and RNA with BrU and testing their close proximity by PLA demonstrates that RNA synthesis in individual cells resumes about 30–45 mi...

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Autores principales: Petruk, Svetlana, Fenstermaker, Tyler K., Black, Kathryn L., Brock, Hugh W., Mazo, Alexander
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4891662/
https://www.ncbi.nlm.nih.gov/pubmed/27256324
http://dx.doi.org/10.1038/srep27313
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author Petruk, Svetlana
Fenstermaker, Tyler K.
Black, Kathryn L.
Brock, Hugh W.
Mazo, Alexander
author_facet Petruk, Svetlana
Fenstermaker, Tyler K.
Black, Kathryn L.
Brock, Hugh W.
Mazo, Alexander
author_sort Petruk, Svetlana
collection PubMed
description We describe a proximity ligation assay (PLA)-based method of assessing association of DNA and RNA in single cells during the cell cycle. Pulse-labeling of DNA with EdU and RNA with BrU and testing their close proximity by PLA demonstrates that RNA synthesis in individual cells resumes about 30–45 min after DNA replication. Consistent with this conclusion, RNA Pol II phosphorylated at Ser2 of its CTD is detected at the same time as RNA transcripts on nascent DNA. Our results also show that RNA is associated with DNA foci during all stages of mitosis.
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spelling pubmed-48916622016-06-09 Detection of RNA-DNA association by a proximity ligation-based method Petruk, Svetlana Fenstermaker, Tyler K. Black, Kathryn L. Brock, Hugh W. Mazo, Alexander Sci Rep Article We describe a proximity ligation assay (PLA)-based method of assessing association of DNA and RNA in single cells during the cell cycle. Pulse-labeling of DNA with EdU and RNA with BrU and testing their close proximity by PLA demonstrates that RNA synthesis in individual cells resumes about 30–45 min after DNA replication. Consistent with this conclusion, RNA Pol II phosphorylated at Ser2 of its CTD is detected at the same time as RNA transcripts on nascent DNA. Our results also show that RNA is associated with DNA foci during all stages of mitosis. Nature Publishing Group 2016-06-03 /pmc/articles/PMC4891662/ /pubmed/27256324 http://dx.doi.org/10.1038/srep27313 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Petruk, Svetlana
Fenstermaker, Tyler K.
Black, Kathryn L.
Brock, Hugh W.
Mazo, Alexander
Detection of RNA-DNA association by a proximity ligation-based method
title Detection of RNA-DNA association by a proximity ligation-based method
title_full Detection of RNA-DNA association by a proximity ligation-based method
title_fullStr Detection of RNA-DNA association by a proximity ligation-based method
title_full_unstemmed Detection of RNA-DNA association by a proximity ligation-based method
title_short Detection of RNA-DNA association by a proximity ligation-based method
title_sort detection of rna-dna association by a proximity ligation-based method
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4891662/
https://www.ncbi.nlm.nih.gov/pubmed/27256324
http://dx.doi.org/10.1038/srep27313
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