Cargando…
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...
Autores principales: | , , , , , , , , , |
---|---|
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 |
_version_ | 1783268693547941888 |
---|---|
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. |
format | Online Article Text |
id | pubmed-5627313 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | The Biophysical Society |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT germierthomas realtimeimagingofasinglegenerevealstranscriptioninitiatedlocalconfinement AT kocanovasilvia realtimeimagingofasinglegenerevealstranscriptioninitiatedlocalconfinement AT walthernike realtimeimagingofasinglegenerevealstranscriptioninitiatedlocalconfinement AT bancaudaurelien realtimeimagingofasinglegenerevealstranscriptioninitiatedlocalconfinement AT shabanhaithamahmed realtimeimagingofasinglegenerevealstranscriptioninitiatedlocalconfinement AT sellouhafida realtimeimagingofasinglegenerevealstranscriptioninitiatedlocalconfinement AT politiantoniozaccaria realtimeimagingofasinglegenerevealstranscriptioninitiatedlocalconfinement AT ellenbergjan realtimeimagingofasinglegenerevealstranscriptioninitiatedlocalconfinement AT gallardofranck realtimeimagingofasinglegenerevealstranscriptioninitiatedlocalconfinement AT bystrickykerstin realtimeimagingofasinglegenerevealstranscriptioninitiatedlocalconfinement |