Formation of nuclear condensates by the Mediator complex subunit Med15 in mammalian cells

Abstract Background The Mediator complex is an evolutionarily conserved multi-subunit protein complex that plays major roles in transcriptional activation and is essential for cell growth, proliferation, and differentiation. Recent studies revealed that some Mediator subunits formed nuclear condensa...

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Autores principales:	Yuanyuan Shi, Jian Chen, Wei-Jie Zeng, Miao Li, Wenxue Zhao, Xing-Ding Zhang, Jie Yao
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Nuclear condensates Mediator Med15 Transcription Cell imaging Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/2ed5b116a42a4485b6a610fadcc9822f
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spelling	oai:doaj.org-article:2ed5b116a42a4485b6a610fadcc9822f2021-11-21T12:42:23ZFormation of nuclear condensates by the Mediator complex subunit Med15 in mammalian cells10.1186/s12915-021-01178-y1741-7007https://doaj.org/article/2ed5b116a42a4485b6a610fadcc9822f2021-11-01T00:00:00Zhttps://doi.org/10.1186/s12915-021-01178-yhttps://doaj.org/toc/1741-7007Abstract Background The Mediator complex is an evolutionarily conserved multi-subunit protein complex that plays major roles in transcriptional activation and is essential for cell growth, proliferation, and differentiation. Recent studies revealed that some Mediator subunits formed nuclear condensates that may facilitate enhancer-promoter interactions and gene activation. The assembly, regulation, and functions of these nuclear condensates remain to be further understood. Results We found that Med15, a subunit in the tail module of the Mediator complex, formed nuclear condensates through a novel mechanism. Nuclear foci of Med15 were detected by both immunostaining of endogenous proteins and live cell imaging. Like Med1 foci and many other biomolecular condensates, Med15 foci were sensitive to 1, 6-Hexanediol and showed rapid recovery during fluorescence recovery after photobleaching. Interestingly, overexpressing DYRK3, a dual-specificity kinase that controls the phase transition of membraneless organelles, appeared to disrupt Med1 foci and Med15 foci. We identified two regions that are required to form Med15 nuclear condensates: the glutamine-rich intrinsically disordered region (IDR) and a short downstream hydrophobic motif. The optodroplet assay revealed that both the IDR and the C-terminal region of Med15 contributed to intracellular phase separation. Conclusions We identified that the Mediator complex subunit Med15 formed nuclear condensates and characterized their features in living cells. Our work suggests that Med15 plays a role in the assembly of transcription coactivator condensates in the nucleus and identifies Med15 regions that contribute to phase separation.Yuanyuan ShiJian ChenWei-Jie ZengMiao LiWenxue ZhaoXing-Ding ZhangJie YaoBMCarticleNuclear condensatesMediatorMed15TranscriptionCell imagingBiology (General)QH301-705.5ENBMC Biology, Vol 19, Iss 1, Pp 1-17 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Nuclear condensates Mediator Med15 Transcription Cell imaging Biology (General) QH301-705.5
spellingShingle	Nuclear condensates Mediator Med15 Transcription Cell imaging Biology (General) QH301-705.5 Yuanyuan Shi Jian Chen Wei-Jie Zeng Miao Li Wenxue Zhao Xing-Ding Zhang Jie Yao Formation of nuclear condensates by the Mediator complex subunit Med15 in mammalian cells
description	Abstract Background The Mediator complex is an evolutionarily conserved multi-subunit protein complex that plays major roles in transcriptional activation and is essential for cell growth, proliferation, and differentiation. Recent studies revealed that some Mediator subunits formed nuclear condensates that may facilitate enhancer-promoter interactions and gene activation. The assembly, regulation, and functions of these nuclear condensates remain to be further understood. Results We found that Med15, a subunit in the tail module of the Mediator complex, formed nuclear condensates through a novel mechanism. Nuclear foci of Med15 were detected by both immunostaining of endogenous proteins and live cell imaging. Like Med1 foci and many other biomolecular condensates, Med15 foci were sensitive to 1, 6-Hexanediol and showed rapid recovery during fluorescence recovery after photobleaching. Interestingly, overexpressing DYRK3, a dual-specificity kinase that controls the phase transition of membraneless organelles, appeared to disrupt Med1 foci and Med15 foci. We identified two regions that are required to form Med15 nuclear condensates: the glutamine-rich intrinsically disordered region (IDR) and a short downstream hydrophobic motif. The optodroplet assay revealed that both the IDR and the C-terminal region of Med15 contributed to intracellular phase separation. Conclusions We identified that the Mediator complex subunit Med15 formed nuclear condensates and characterized their features in living cells. Our work suggests that Med15 plays a role in the assembly of transcription coactivator condensates in the nucleus and identifies Med15 regions that contribute to phase separation.
format	article
author	Yuanyuan Shi Jian Chen Wei-Jie Zeng Miao Li Wenxue Zhao Xing-Ding Zhang Jie Yao
author_facet	Yuanyuan Shi Jian Chen Wei-Jie Zeng Miao Li Wenxue Zhao Xing-Ding Zhang Jie Yao
author_sort	Yuanyuan Shi
title	Formation of nuclear condensates by the Mediator complex subunit Med15 in mammalian cells
title_short	Formation of nuclear condensates by the Mediator complex subunit Med15 in mammalian cells
title_full	Formation of nuclear condensates by the Mediator complex subunit Med15 in mammalian cells
title_fullStr	Formation of nuclear condensates by the Mediator complex subunit Med15 in mammalian cells
title_full_unstemmed	Formation of nuclear condensates by the Mediator complex subunit Med15 in mammalian cells
title_sort	formation of nuclear condensates by the mediator complex subunit med15 in mammalian cells
publisher	BMC
publishDate	2021
url	https://doaj.org/article/2ed5b116a42a4485b6a610fadcc9822f
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Formation of nuclear condensates by the Mediator complex subunit Med15 in mammalian cells

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