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Real-Time Imaging of a Single Gene Reveals Transcription-Initiated Local Confinement

Genome dynamics are intimately linked to the regulation of gene expression, the most fundamental mechanism in biology, yet we still do not know whether the very process of transcription drives spatial organization at specific gene loci. Here, we have optimized the ANCHOR/ParB DNA-labeling system for...

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Autores principales: Germier, Thomas, Kocanova, Silvia, Walther, Nike, Bancaud, Aurélien, Shaban, Haitham Ahmed, Sellou, Hafida, Politi, Antonio Zaccaria, Ellenberg, Jan, Gallardo, Franck, Bystricky, Kerstin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Biophysical Society 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5627313/
https://www.ncbi.nlm.nih.gov/pubmed/28978433
http://dx.doi.org/10.1016/j.bpj.2017.08.014
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author Germier, Thomas
Kocanova, Silvia
Walther, Nike
Bancaud, Aurélien
Shaban, Haitham Ahmed
Sellou, Hafida
Politi, Antonio Zaccaria
Ellenberg, Jan
Gallardo, Franck
Bystricky, Kerstin
author_facet Germier, Thomas
Kocanova, Silvia
Walther, Nike
Bancaud, Aurélien
Shaban, Haitham Ahmed
Sellou, Hafida
Politi, Antonio Zaccaria
Ellenberg, Jan
Gallardo, Franck
Bystricky, Kerstin
author_sort Germier, Thomas
collection PubMed
description Genome dynamics are intimately linked to the regulation of gene expression, the most fundamental mechanism in biology, yet we still do not know whether the very process of transcription drives spatial organization at specific gene loci. Here, we have optimized the ANCHOR/ParB DNA-labeling system for real-time imaging of a single-copy, estrogen-inducible transgene in human cells. Motion of an ANCHOR3-tagged DNA locus was recorded in the same cell before and during the appearance of nascent MS2-labeled mRNA. We found that transcription initiation by RNA polymerase 2 resulted in confinement of the mRNA-producing gene domain within minutes. Transcription-induced confinement occurred in each single cell independently of initial, highly heterogeneous mobility. Constrained mobility was maintained even when inhibiting polymerase elongation. Chromatin motion at constant step size within a largely confined area hence leads to increased collisions that are compatible with the formation of gene-specific chromatin domains, and reflect the assembly of functional protein hubs and DNA processing during the rate-limiting steps of transcription.
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spelling pubmed-56273132018-10-03 Real-Time Imaging of a Single Gene Reveals Transcription-Initiated Local Confinement Germier, Thomas Kocanova, Silvia Walther, Nike Bancaud, Aurélien Shaban, Haitham Ahmed Sellou, Hafida Politi, Antonio Zaccaria Ellenberg, Jan Gallardo, Franck Bystricky, Kerstin Biophys J Nucleic Acids and Genome Biophysics Genome dynamics are intimately linked to the regulation of gene expression, the most fundamental mechanism in biology, yet we still do not know whether the very process of transcription drives spatial organization at specific gene loci. Here, we have optimized the ANCHOR/ParB DNA-labeling system for real-time imaging of a single-copy, estrogen-inducible transgene in human cells. Motion of an ANCHOR3-tagged DNA locus was recorded in the same cell before and during the appearance of nascent MS2-labeled mRNA. We found that transcription initiation by RNA polymerase 2 resulted in confinement of the mRNA-producing gene domain within minutes. Transcription-induced confinement occurred in each single cell independently of initial, highly heterogeneous mobility. Constrained mobility was maintained even when inhibiting polymerase elongation. Chromatin motion at constant step size within a largely confined area hence leads to increased collisions that are compatible with the formation of gene-specific chromatin domains, and reflect the assembly of functional protein hubs and DNA processing during the rate-limiting steps of transcription. The Biophysical Society 2017-10-03 2017-10-03 /pmc/articles/PMC5627313/ /pubmed/28978433 http://dx.doi.org/10.1016/j.bpj.2017.08.014 Text en © 2017 Biophysical Society. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Nucleic Acids and Genome Biophysics
Germier, Thomas
Kocanova, Silvia
Walther, Nike
Bancaud, Aurélien
Shaban, Haitham Ahmed
Sellou, Hafida
Politi, Antonio Zaccaria
Ellenberg, Jan
Gallardo, Franck
Bystricky, Kerstin
Real-Time Imaging of a Single Gene Reveals Transcription-Initiated Local Confinement
title Real-Time Imaging of a Single Gene Reveals Transcription-Initiated Local Confinement
title_full Real-Time Imaging of a Single Gene Reveals Transcription-Initiated Local Confinement
title_fullStr Real-Time Imaging of a Single Gene Reveals Transcription-Initiated Local Confinement
title_full_unstemmed Real-Time Imaging of a Single Gene Reveals Transcription-Initiated Local Confinement
title_short Real-Time Imaging of a Single Gene Reveals Transcription-Initiated Local Confinement
title_sort real-time imaging of a single gene reveals transcription-initiated local confinement
topic Nucleic Acids and Genome Biophysics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5627313/
https://www.ncbi.nlm.nih.gov/pubmed/28978433
http://dx.doi.org/10.1016/j.bpj.2017.08.014
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