Cargando…
Differentiating cancer cells reveal early large-scale genome regulation by pericentric domains
Finding out how cells prepare for fate change during differentiation commitment was our task. To address whether the constitutive pericentromere-associated domains (PADs) may be involved, we used a model system with known transcriptome data, MCF-7 breast cancer cells treated with the ErbB3 ligand he...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Biophysical Society
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7896032/ https://www.ncbi.nlm.nih.gov/pubmed/33453273 http://dx.doi.org/10.1016/j.bpj.2021.01.002 |
_version_ | 1783653472045891584 |
---|---|
author | Krigerts, Jekabs Salmina, Kristine Freivalds, Talivaldis Zayakin, Pawel Rumnieks, Felikss Inashkina, Inna Giuliani, Alessandro Hausmann, Michael Erenpreisa, Jekaterina |
author_facet | Krigerts, Jekabs Salmina, Kristine Freivalds, Talivaldis Zayakin, Pawel Rumnieks, Felikss Inashkina, Inna Giuliani, Alessandro Hausmann, Michael Erenpreisa, Jekaterina |
author_sort | Krigerts, Jekabs |
collection | PubMed |
description | Finding out how cells prepare for fate change during differentiation commitment was our task. To address whether the constitutive pericentromere-associated domains (PADs) may be involved, we used a model system with known transcriptome data, MCF-7 breast cancer cells treated with the ErbB3 ligand heregulin (HRG), which induces differentiation and is used in the therapy of cancer. PAD-repressive heterochromatin (H3K9me3), centromere-associated-protein-specific, and active euchromatin (H3K4me3) antibodies, real-time PCR, acridine orange DNA structural test (AOT), and microscopic image analysis were applied. We found a two-step DNA unfolding after 15–20 and 60 min of HRG treatment, respectively. This behavior was consistent with biphasic activation of the early response genes (c-fos - fosL1/myc) and the timing of two transcriptome avalanches reported in the literature. In control, the average number of PADs negatively correlated with their size by scale-free distribution, and centromere clustering in turn correlated with PAD size, both indicating that PADs may create and modulate a suprachromosomal network by fusing and splitting a constant proportion of the constitutive heterochromatin. By 15 min of HRG treatment, the bursting unraveling of PADs from the nucleolus boundary occurred, coinciding with the first step of H3K4me3 chromatin unfolding, confirmed by AOT. The second step after 60 min of HRG treatment was associated with transcription of long noncoding RNA from PADs and peaking of fosL1/c-myc response. We hypothesize that the bursting of PAD clusters under a critical silencing threshold pushes the first transcription avalanche, whereas the destruction of the PAD network enables genome rewiring needed for differentiation repatterning, mediated by early response bivalent genes. |
format | Online Article Text |
id | pubmed-7896032 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | The Biophysical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-78960322022-02-16 Differentiating cancer cells reveal early large-scale genome regulation by pericentric domains Krigerts, Jekabs Salmina, Kristine Freivalds, Talivaldis Zayakin, Pawel Rumnieks, Felikss Inashkina, Inna Giuliani, Alessandro Hausmann, Michael Erenpreisa, Jekaterina Biophys J Articles Finding out how cells prepare for fate change during differentiation commitment was our task. To address whether the constitutive pericentromere-associated domains (PADs) may be involved, we used a model system with known transcriptome data, MCF-7 breast cancer cells treated with the ErbB3 ligand heregulin (HRG), which induces differentiation and is used in the therapy of cancer. PAD-repressive heterochromatin (H3K9me3), centromere-associated-protein-specific, and active euchromatin (H3K4me3) antibodies, real-time PCR, acridine orange DNA structural test (AOT), and microscopic image analysis were applied. We found a two-step DNA unfolding after 15–20 and 60 min of HRG treatment, respectively. This behavior was consistent with biphasic activation of the early response genes (c-fos - fosL1/myc) and the timing of two transcriptome avalanches reported in the literature. In control, the average number of PADs negatively correlated with their size by scale-free distribution, and centromere clustering in turn correlated with PAD size, both indicating that PADs may create and modulate a suprachromosomal network by fusing and splitting a constant proportion of the constitutive heterochromatin. By 15 min of HRG treatment, the bursting unraveling of PADs from the nucleolus boundary occurred, coinciding with the first step of H3K4me3 chromatin unfolding, confirmed by AOT. The second step after 60 min of HRG treatment was associated with transcription of long noncoding RNA from PADs and peaking of fosL1/c-myc response. We hypothesize that the bursting of PAD clusters under a critical silencing threshold pushes the first transcription avalanche, whereas the destruction of the PAD network enables genome rewiring needed for differentiation repatterning, mediated by early response bivalent genes. The Biophysical Society 2021-02-16 2021-01-14 /pmc/articles/PMC7896032/ /pubmed/33453273 http://dx.doi.org/10.1016/j.bpj.2021.01.002 Text en © 2021 Biophysical Society. http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Articles Krigerts, Jekabs Salmina, Kristine Freivalds, Talivaldis Zayakin, Pawel Rumnieks, Felikss Inashkina, Inna Giuliani, Alessandro Hausmann, Michael Erenpreisa, Jekaterina Differentiating cancer cells reveal early large-scale genome regulation by pericentric domains |
title | Differentiating cancer cells reveal early large-scale genome regulation by pericentric domains |
title_full | Differentiating cancer cells reveal early large-scale genome regulation by pericentric domains |
title_fullStr | Differentiating cancer cells reveal early large-scale genome regulation by pericentric domains |
title_full_unstemmed | Differentiating cancer cells reveal early large-scale genome regulation by pericentric domains |
title_short | Differentiating cancer cells reveal early large-scale genome regulation by pericentric domains |
title_sort | differentiating cancer cells reveal early large-scale genome regulation by pericentric domains |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7896032/ https://www.ncbi.nlm.nih.gov/pubmed/33453273 http://dx.doi.org/10.1016/j.bpj.2021.01.002 |
work_keys_str_mv | AT krigertsjekabs differentiatingcancercellsrevealearlylargescalegenomeregulationbypericentricdomains AT salminakristine differentiatingcancercellsrevealearlylargescalegenomeregulationbypericentricdomains AT freivaldstalivaldis differentiatingcancercellsrevealearlylargescalegenomeregulationbypericentricdomains AT zayakinpawel differentiatingcancercellsrevealearlylargescalegenomeregulationbypericentricdomains AT rumnieksfelikss differentiatingcancercellsrevealearlylargescalegenomeregulationbypericentricdomains AT inashkinainna differentiatingcancercellsrevealearlylargescalegenomeregulationbypericentricdomains AT giulianialessandro differentiatingcancercellsrevealearlylargescalegenomeregulationbypericentricdomains AT hausmannmichael differentiatingcancercellsrevealearlylargescalegenomeregulationbypericentricdomains AT erenpreisajekaterina differentiatingcancercellsrevealearlylargescalegenomeregulationbypericentricdomains |