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RNA polymerase II condensate formation and association with Cajal and histone locus bodies in living human cells

View ORCID ProfileTakashi Imada, View ORCID ProfileTakeshi Shimi, Ai Kaiho, View ORCID ProfileYasushi Saeki, View ORCID ProfileHiroshi Kimura
doi: https://doi.org/10.1101/2020.12.25.424380
Takashi Imada
1Graduate School of Life Science, Tokyo Institute of Technology, Yokohama, Japan
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Takeshi Shimi
2World Research Hub Initiative, Institute of Innovative Research, Tokyo Institute of Technology, Yokohama, Japan
3Cell Biology Center, Institute of Innovative Research, Tokyo Institute of Technology, Yokohama, Japan
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Ai Kaiho
4Protein Metabolism Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan
5Institute for Advanced Life Sciences, Hoshi University, Tokyo, Japan
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Yasushi Saeki
5Institute for Advanced Life Sciences, Hoshi University, Tokyo, Japan
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Hiroshi Kimura
1Graduate School of Life Science, Tokyo Institute of Technology, Yokohama, Japan
2World Research Hub Initiative, Institute of Innovative Research, Tokyo Institute of Technology, Yokohama, Japan
3Cell Biology Center, Institute of Innovative Research, Tokyo Institute of Technology, Yokohama, Japan
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  • For correspondence: hkimura@bio.titech.ac.jp
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ABSTRACT

In eukaryotic nuclei, a number of phase-separated nuclear bodies (NBs) are present. RNA polymerase II (Pol II) is the main player in transcription and forms large condensates in addition to localizing at numerous transcription foci. Cajal bodies (CBs) and histone locus bodies (HLBs) are NBs that are involved in transcriptional and post-transcriptional regulation of small nuclear RNA and histone genes. By live-cell imaging using human HCT116 cells, we here show that Pol II condensates (PCs) nucleated near CBs and HLBs, and the number of PCs increased during S phase concomitantly with the activation period of histone genes. Ternary PC–CB– HLB associates were formed via three pathways: nucleation of PCs and HLBs near CBs, interaction between preformed PC–HLBs with CBs, and nucleation of PCs near preformed CB– HLBs. Coilin knockout increased the co-localization rate between PCs and HLBs, whereas the number, nucleation timing, and phosphorylation status of PCs remained unchanged. Depletion of PCs did not affect CBs and HLBs. Treatment with 1,6-hexanediol revealed that PCs were more liquid-like than CBs and HLBs. Thus, PCs are dynamic structures often nucleated following the activation of gene clusters associated with other NBs. (187 words)

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC 4.0 International license.
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Posted December 25, 2020.
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RNA polymerase II condensate formation and association with Cajal and histone locus bodies in living human cells
Takashi Imada, Takeshi Shimi, Ai Kaiho, Yasushi Saeki, Hiroshi Kimura
bioRxiv 2020.12.25.424380; doi: https://doi.org/10.1101/2020.12.25.424380
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RNA polymerase II condensate formation and association with Cajal and histone locus bodies in living human cells
Takashi Imada, Takeshi Shimi, Ai Kaiho, Yasushi Saeki, Hiroshi Kimura
bioRxiv 2020.12.25.424380; doi: https://doi.org/10.1101/2020.12.25.424380

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