STREAMING-tag system reveals spatiotemporal relationships between transcriptional regulatory factors and transcriptional activity

Transcription is a dynamic process. To detect the dynamic relationship among protein clusters of RNA polymerase II and coactivators, gene loci, and transcriptional activity, we insert an MS2 repeat, a TetO repeat, and inteins with a selection marker just downstream of the transcription start site. B...

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Bibliographic Details
Published in:	Nature communications Vol. 13; no. 1; p. 7672
Main Authors:	Ohishi, Hiroaki, Shimada, Seiru, Uchino, Satoshi, Li, Jieru, Sato, Yuko, Shintani, Manabu, Owada, Hitoshi, Ohkawa, Yasuyuki, Pertsinidis, Alexandros, Yamamoto, Takashi, Kimura, Hiroshi, Ochiai, Hiroshi
Format:	Journal Article
Language:	English
Published:	London Nature Publishing Group UK 20-12-2022 Nature Publishing Group Nature Portfolio
Subjects:	13/100 14/19 14/32 14/34 14/35 14/63 38/71 38/88 631/337/572 631/80/2373 Animals Clusters DNA-directed RNA polymerase Embryo cells Gene Expression Regulation Gene regulation Humanities and Social Sciences Inteins Inteins - genetics Mice multidisciplinary Nuclear Proteins - metabolism Proteins Reporter gene Ribonucleic acid RNA RNA polymerase RNA polymerase II RNA Polymerase II - genetics RNA Polymerase II - metabolism Science Science (multidisciplinary) Stem cell transplantation Stem cells Transcription Transcription factors Transcription Factors - genetics Transcription Factors - metabolism Transcription initiation Transcription, Genetic
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Summary:	Transcription is a dynamic process. To detect the dynamic relationship among protein clusters of RNA polymerase II and coactivators, gene loci, and transcriptional activity, we insert an MS2 repeat, a TetO repeat, and inteins with a selection marker just downstream of the transcription start site. By optimizing the individual elements, we develop the Spliced TetO REpeAt, MS2 repeat, and INtein sandwiched reporter Gene tag (STREAMING-tag) system. Clusters of RNA polymerase II and BRD4 are observed proximal to the transcription start site of Nanog when the gene is transcribed in mouse embryonic stem cells. In contrast, clusters of MED19 and MED22 tend to be located near the transcription start site, even without transcription activity. Thus, the STREAMING-tag system reveals the spatiotemporal relationships between transcriptional activity and protein clusters near the gene. This powerful tool is useful for quantitatively understanding transcriptional regulation in living cells. Using the newly developed STREAMING-tag system, the authors find that clusters of RNA polymerase II and BRD4 are formed specifically in the transcriptionally active state near the Nanog gene in mouse embryonic stem cells.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	2041-1723 2041-1723
DOI:	10.1038/s41467-022-35286-2