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Real-time chromatin dynamics at the single gene level during transcription activation

Thomas Germier, Silvia Kocanova, Nike Walther, Aurélien Bancaud, Haitham Ahmed Shaban, Hafida Sellou, Antonio Zaccaria Politi, Jan Ellenberg, Franck Gallardo, Kerstin Bystricky
doi: https://doi.org/10.1101/111179
Thomas Germier
1Laboratoire de Biologie Moléculaire Eucaryote (LBME); Centre de Biologie Intégrative (CBI); Université de Toulouse; CNRS; UPS; 31062 Toulouse, France
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Silvia Kocanova
1Laboratoire de Biologie Moléculaire Eucaryote (LBME); Centre de Biologie Intégrative (CBI); Université de Toulouse; CNRS; UPS; 31062 Toulouse, France
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Nike Walther
2European Molecular Biology Laboratory (EMBL); Heidelberg, Germany
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Aurélien Bancaud
3Laboratoire de Automatismes et Architecture des Systèmes(LAAS); CNRS; UPS; 31000 Toulouse, France
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Haitham Ahmed Shaban
1Laboratoire de Biologie Moléculaire Eucaryote (LBME); Centre de Biologie Intégrative (CBI); Université de Toulouse; CNRS; UPS; 31062 Toulouse, France
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Hafida Sellou
1Laboratoire de Biologie Moléculaire Eucaryote (LBME); Centre de Biologie Intégrative (CBI); Université de Toulouse; CNRS; UPS; 31062 Toulouse, France
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Antonio Zaccaria Politi
2European Molecular Biology Laboratory (EMBL); Heidelberg, Germany
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Jan Ellenberg
2European Molecular Biology Laboratory (EMBL); Heidelberg, Germany
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Franck Gallardo
1Laboratoire de Biologie Moléculaire Eucaryote (LBME); Centre de Biologie Intégrative (CBI); Université de Toulouse; CNRS; UPS; 31062 Toulouse, France
4Institut des Technologies Avancées du Vivant (ITAV); Universite de Toulouse; CNRS; UPS; INSA; 31000 Toulouse, France
5NeoVirTech S.A.; 31000 Toulouse, France
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Kerstin Bystricky
1Laboratoire de Biologie Moléculaire Eucaryote (LBME); Centre de Biologie Intégrative (CBI); Université de Toulouse; CNRS; UPS; 31062 Toulouse, France
4Institut des Technologies Avancées du Vivant (ITAV); Universite de Toulouse; CNRS; UPS; INSA; 31000 Toulouse, France
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Abstract

Genome dynamics relate to regulation of gene expression, the most fundamental process in biology. Yet we still do not know whether the very process of transcription drives spatial organization and chromatin conformation at specific gene loci. To address this issue, we have optimized the ANCHOR/ParB DNA labeling system for real-time imaging and quantitative analysis of the dynamics of a single-copy transgene in human cells. Transcription of the transgene under the control of the endogenous Cyclin D1 promoter was induced by addition of 17β-estradiol. Motion of the ANCHOR3-tagged DNA locus was recorded in the same cell prior to and during appearance of nascent mRNA visualized using the MS2 system. We found that transcription initiation resulted in rapid confinement of the mRNA-producing gene. The confinement was maintained even upon inhibition of pol2 elongation. It did not occur when recruitment of pol2 or transcription initiation was blocked by anti-estrogens or Triptolide. These results suggest that preinitiation complex formation and concomitant reorganization of the chromatin domain constrains freedom of movement of an induced gene’s promoter within minutes. Confined diffusion reflects assembly of functional protein hubs and DNA processing during the rate-limiting steps of transcription.

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Posted February 23, 2017.
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Real-time chromatin dynamics at the single gene level during transcription activation
Thomas Germier, Silvia Kocanova, Nike Walther, Aurélien Bancaud, Haitham Ahmed Shaban, Hafida Sellou, Antonio Zaccaria Politi, Jan Ellenberg, Franck Gallardo, Kerstin Bystricky
bioRxiv 111179; doi: https://doi.org/10.1101/111179
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Real-time chromatin dynamics at the single gene level during transcription activation
Thomas Germier, Silvia Kocanova, Nike Walther, Aurélien Bancaud, Haitham Ahmed Shaban, Hafida Sellou, Antonio Zaccaria Politi, Jan Ellenberg, Franck Gallardo, Kerstin Bystricky
bioRxiv 111179; doi: https://doi.org/10.1101/111179

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