Cargando…

Proximity-dependent biotin labelling reveals CP190 as an EcR/Usp molecular partner

Proximity-dependent biotin labelling revealed undescribed participants of the ecdysone response in Drosophila. Two labelling enzymes (BioID2 and APEX2) were fused to EcR or Usp to biotin label the surrounding proteins. The EcR/Usp heterodimer was found to collaborate with nuclear pore subunits, chro...

Descripción completa

Detalles Bibliográficos
Autores principales: Mazina, Marina Yu., Ziganshin, Rustam H., Magnitov, Mikhail D., Golovnin, Anton K., Vorobyeva, Nadezhda E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7075897/
https://www.ncbi.nlm.nih.gov/pubmed/32179799
http://dx.doi.org/10.1038/s41598-020-61514-0
_version_ 1783507110726729728
author Mazina, Marina Yu.
Ziganshin, Rustam H.
Magnitov, Mikhail D.
Golovnin, Anton K.
Vorobyeva, Nadezhda E.
author_facet Mazina, Marina Yu.
Ziganshin, Rustam H.
Magnitov, Mikhail D.
Golovnin, Anton K.
Vorobyeva, Nadezhda E.
author_sort Mazina, Marina Yu.
collection PubMed
description Proximity-dependent biotin labelling revealed undescribed participants of the ecdysone response in Drosophila. Two labelling enzymes (BioID2 and APEX2) were fused to EcR or Usp to biotin label the surrounding proteins. The EcR/Usp heterodimer was found to collaborate with nuclear pore subunits, chromatin remodelers, and architectural proteins. Many proteins identified through proximity-dependent labelling with EcR/Usp were described previously as functional components of an ecdysone response, corroborating the potency of this labelling method. A link to ecdysone response was confirmed for some newly discovered regulators by immunoprecipitation of prepupal nuclear extract with anti-EcR antibodies and functional experiments in Drosophila S2 cells. A more in-depth study was conducted to clarify the association of EcR/Usp with one of the detected proteins, CP190, a well-described cofactor of Drosophila insulators. CP190 was found to co-immunoprecipitate with the EcR subunit of EcR/Usp in a 20E-independent manner. ChIP-Seq experiments revealed only partial overlapping between CP190 and EcR bound sites in the Drosophila genome and complete absence of CP190 binding at 20E-dependent enhancers. Analysis of Hi-C data demonstrated an existence of remote interactions between 20E-dependent enhancers and CP190 sites which suggests formation of a protein complex between EcR/Usp and CP190 through the space. Our results support the previous concept that CP190 has a role in stabilization of specific chromatin loops for proper activation of transcription of genes regulated by 20E hormone.
format Online
Article
Text
id pubmed-7075897
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-70758972020-03-23 Proximity-dependent biotin labelling reveals CP190 as an EcR/Usp molecular partner Mazina, Marina Yu. Ziganshin, Rustam H. Magnitov, Mikhail D. Golovnin, Anton K. Vorobyeva, Nadezhda E. Sci Rep Article Proximity-dependent biotin labelling revealed undescribed participants of the ecdysone response in Drosophila. Two labelling enzymes (BioID2 and APEX2) were fused to EcR or Usp to biotin label the surrounding proteins. The EcR/Usp heterodimer was found to collaborate with nuclear pore subunits, chromatin remodelers, and architectural proteins. Many proteins identified through proximity-dependent labelling with EcR/Usp were described previously as functional components of an ecdysone response, corroborating the potency of this labelling method. A link to ecdysone response was confirmed for some newly discovered regulators by immunoprecipitation of prepupal nuclear extract with anti-EcR antibodies and functional experiments in Drosophila S2 cells. A more in-depth study was conducted to clarify the association of EcR/Usp with one of the detected proteins, CP190, a well-described cofactor of Drosophila insulators. CP190 was found to co-immunoprecipitate with the EcR subunit of EcR/Usp in a 20E-independent manner. ChIP-Seq experiments revealed only partial overlapping between CP190 and EcR bound sites in the Drosophila genome and complete absence of CP190 binding at 20E-dependent enhancers. Analysis of Hi-C data demonstrated an existence of remote interactions between 20E-dependent enhancers and CP190 sites which suggests formation of a protein complex between EcR/Usp and CP190 through the space. Our results support the previous concept that CP190 has a role in stabilization of specific chromatin loops for proper activation of transcription of genes regulated by 20E hormone. Nature Publishing Group UK 2020-03-16 /pmc/articles/PMC7075897/ /pubmed/32179799 http://dx.doi.org/10.1038/s41598-020-61514-0 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Mazina, Marina Yu.
Ziganshin, Rustam H.
Magnitov, Mikhail D.
Golovnin, Anton K.
Vorobyeva, Nadezhda E.
Proximity-dependent biotin labelling reveals CP190 as an EcR/Usp molecular partner
title Proximity-dependent biotin labelling reveals CP190 as an EcR/Usp molecular partner
title_full Proximity-dependent biotin labelling reveals CP190 as an EcR/Usp molecular partner
title_fullStr Proximity-dependent biotin labelling reveals CP190 as an EcR/Usp molecular partner
title_full_unstemmed Proximity-dependent biotin labelling reveals CP190 as an EcR/Usp molecular partner
title_short Proximity-dependent biotin labelling reveals CP190 as an EcR/Usp molecular partner
title_sort proximity-dependent biotin labelling reveals cp190 as an ecr/usp molecular partner
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7075897/
https://www.ncbi.nlm.nih.gov/pubmed/32179799
http://dx.doi.org/10.1038/s41598-020-61514-0
work_keys_str_mv AT mazinamarinayu proximitydependentbiotinlabellingrevealscp190asanecruspmolecularpartner
AT ziganshinrustamh proximitydependentbiotinlabellingrevealscp190asanecruspmolecularpartner
AT magnitovmikhaild proximitydependentbiotinlabellingrevealscp190asanecruspmolecularpartner
AT golovninantonk proximitydependentbiotinlabellingrevealscp190asanecruspmolecularpartner
AT vorobyevanadezhdae proximitydependentbiotinlabellingrevealscp190asanecruspmolecularpartner