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Replisome loading reduces chromatin motion independent of DNA synthesis

Pabba, Maruthi Kumar ; Ritter, Christian ; Chagin, Vadim O. ; Meyer, Janis ; Celikay, Kerem ; Stear, Jeffrey H. ; Loerke, Dinah ; Kolobynina, Ksenia ; Prorok, Paulina ; Schmid, Alice Kristin ; Leonhardt, Heinrich ; Rohr, Karl ; Cardoso, M. Cristina (2023)
Replisome loading reduces chromatin motion independent of DNA synthesis.
In: eLife, 12
doi: 10.7554/eLife.87572
Article, Bibliographie

Abstract

Chromatin has been shown to undergo diffusional motion, which is affected during gene transcription by RNA polymerase activity. However, the relationship between chromatin mobility and other genomic processes remains unclear. Hence, we set out to label the DNA directly in a sequence unbiased manner and followed labeled chromatin dynamics in interphase human cells expressing GFP-tagged proliferating cell nuclear antigen (PCNA), a cell cycle marker and core component of the DNA replication machinery. We detected decreased chromatin mobility during the S-phase compared to G1 and G2 phases in tumor as well as normal diploid cells using automated particle tracking. To gain insight into the dynamical organization of the genome during DNA replication, we determined labeled chromatin domain sizes and analyzed their motion in replicating cells. By correlating chromatin mobility proximal to the active sites of DNA synthesis, we showed that chromatin motion was locally constrained at the sites of DNA replication. Furthermore, inhibiting DNA synthesis led to increased loading of DNA polymerases. This was accompanied by accumulation of the single-stranded DNA binding protein on the chromatin and activation of DNA helicases further restricting local chromatin motion. We, therefore, propose that it is the loading of replisomes but not their catalytic activity that reduces the dynamics of replicating chromatin segments in the S-phase as well as their accessibility and probability of interactions with other genomic regions.

Item Type: Article
Erschienen: 2023
Creators: Pabba, Maruthi Kumar ; Ritter, Christian ; Chagin, Vadim O. ; Meyer, Janis ; Celikay, Kerem ; Stear, Jeffrey H. ; Loerke, Dinah ; Kolobynina, Ksenia ; Prorok, Paulina ; Schmid, Alice Kristin ; Leonhardt, Heinrich ; Rohr, Karl ; Cardoso, M. Cristina
Type of entry: Bibliographie
Title: Replisome loading reduces chromatin motion independent of DNA synthesis
Language: English
Date: 31 October 2023
Publisher: eLife Sciences Publications
Journal or Publication Title: eLife
Volume of the journal: 12
DOI: 10.7554/eLife.87572
Abstract:

Chromatin has been shown to undergo diffusional motion, which is affected during gene transcription by RNA polymerase activity. However, the relationship between chromatin mobility and other genomic processes remains unclear. Hence, we set out to label the DNA directly in a sequence unbiased manner and followed labeled chromatin dynamics in interphase human cells expressing GFP-tagged proliferating cell nuclear antigen (PCNA), a cell cycle marker and core component of the DNA replication machinery. We detected decreased chromatin mobility during the S-phase compared to G1 and G2 phases in tumor as well as normal diploid cells using automated particle tracking. To gain insight into the dynamical organization of the genome during DNA replication, we determined labeled chromatin domain sizes and analyzed their motion in replicating cells. By correlating chromatin mobility proximal to the active sites of DNA synthesis, we showed that chromatin motion was locally constrained at the sites of DNA replication. Furthermore, inhibiting DNA synthesis led to increased loading of DNA polymerases. This was accompanied by accumulation of the single-stranded DNA binding protein on the chromatin and activation of DNA helicases further restricting local chromatin motion. We, therefore, propose that it is the loading of replisomes but not their catalytic activity that reduces the dynamics of replicating chromatin segments in the S-phase as well as their accessibility and probability of interactions with other genomic regions.

Identification Number: pmid:37906089
Additional Information:

Artikel-ID: RP8757

Divisions: 10 Department of Biology
10 Department of Biology > Cell Biology and Epigenetics
Date Deposited: 07 Nov 2023 08:14
Last Modified: 07 Nov 2023 09:25
PPN: 512983437
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