Perspective Article: Space-time dynamics of genome replication studied with super-resolved microscopy DOI Open Access
Márton Gelléri,

Michael Sterr,

Hilmar Strickfaden

et al.

Postępy Biochemii, Journal Year: 2024, Volume and Issue: 70(1), P. 8 - 21

Published: April 23, 2024

Genome replication requires duplication of the complete set DNA sequences together with nucleosomes and epigenetic signatures. Notwithstanding profound knowledge on mechanistic details replication, major problems genome have remained unresolved. In this perspective article, we consider accessibility machines to all in due course, maintenance functionally important positional structural features chromatid domains during rapid transition CTs into prophase chromosomes two chromatids. We illustrate problem EdU pulse-labeling (10 min) chase experiments (80 performed mouse myeloblast cells. Following light optical serial sectioning nuclei 3D structured illumination microscopy (SIM), seven intensity classes were distinguished as proxies for increasing compaction. cells fixed immediately after pulse-label, observed a relative under-representation EdU-labeled low density classes, representing active nuclear compartment (ANC), an over-representation high inactive (INC). Cells revealed even more pronounced shift classes. This finding contrasts previous studies transcriptional topography demonstrating signatures chromatin RNAPII their discuss these findings current models viewing CDs either frameworks or phase-separated droplets, well methodological limitations that currently prevent integration contrasting evidence spatial transcription common framework dynamic architecture.

Language: Английский

Space-time dynamics of genome replication studied with super-resolved microscopy DOI Creative Commons
Márton Gelléri,

Michael Sterr,

Hilmar Strickfaden

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: March 22, 2024

Abstract Genome replication requires duplication of the complete set DNA sequences together with nucleosomes and epigenetic signatures. Notwithstanding profound knowledge on mechanistic details replication, major problems genome have remained unresolved. In this perspective article, we consider accessibility machines to all in due course, maintenance functionally important positional structural features chromatid domains during rapid transition CTs into prophase chromosomes two chromatids. We illustrate problem EdU pulse-labeling (10 min) chase experiments (80 performed mouse myeloblast cells. Following light optical serial sectioning nuclei 3D structured illumination microscopy (SIM), seven intensity classes were distinguished as proxies for increasing compaction. cells fixed immediately after pulse-label, observed a relative under-representation EdU-labeled low density classes, representing active nuclear compartment (ANC), an over-representation high inactive (INC). Cells revealed even more pronounced shift classes. This finding contrasts previous studies transcriptional topography demonstrating signatures chromatin RNAPII their discuss these findings current models viewing CDs either frameworks or phase-separated droplets, well methodological limitations that currently prevent integration contrasting evidence spatial transcription common framework dynamic architecture.

Language: Английский

Citations

1
Perspective Article: Space-time dynamics of genome replication studied with super-resolved microscopy DOI Open Access
Márton Gelléri,

Michael Sterr,

Hilmar Strickfaden

et al.

Postępy Biochemii, Journal Year: 2024, Volume and Issue: 70(1), P. 8 - 21

Published: April 23, 2024

Genome replication requires duplication of the complete set DNA sequences together with nucleosomes and epigenetic signatures. Notwithstanding profound knowledge on mechanistic details replication, major problems genome have remained unresolved. In this perspective article, we consider accessibility machines to all in due course, maintenance functionally important positional structural features chromatid domains during rapid transition CTs into prophase chromosomes two chromatids. We illustrate problem EdU pulse-labeling (10 min) chase experiments (80 performed mouse myeloblast cells. Following light optical serial sectioning nuclei 3D structured illumination microscopy (SIM), seven intensity classes were distinguished as proxies for increasing compaction. cells fixed immediately after pulse-label, observed a relative under-representation EdU-labeled low density classes, representing active nuclear compartment (ANC), an over-representation high inactive (INC). Cells revealed even more pronounced shift classes. This finding contrasts previous studies transcriptional topography demonstrating signatures chromatin RNAPII their discuss these findings current models viewing CDs either frameworks or phase-separated droplets, well methodological limitations that currently prevent integration contrasting evidence spatial transcription common framework dynamic architecture.

Language: Английский

Citations

0