Single-molecule mapping of replisome progression Journal Article
| Authors: | Claussin, C.; Vazquez, J.; Whitehouse, I. |
| Article Title: | Single-molecule mapping of replisome progression |
| Abstract: | Fundamental aspects of DNA replication, such as the anatomy of replication stall sites, how replisomes are influenced by gene transcription, and whether the progression of sister replisomes is coordinated, are poorly understood. Available techniques do not allow the precise mapping of the positions of individual replisomes on chromatin. We have developed a method called Replicon-seq that entails the excision of full-length replicons by controlled nuclease cleavage at replication forks. Replicons are sequenced using Nanopore, which provides a single-molecule readout of long DNA. Using Replicon-seq, we found that sister replisomes function autonomously and yet progress through chromatin with remarkable consistency. Replication forks that encounter obstacles pause for a short duration but rapidly resume synthesis. The helicase Rrm3 plays a critical role both in mitigating the effect of protein barriers and with facilitating efficient termination. Replicon-seq provides a high-resolution means of defining how individual replisomes move across the genome. © 2022 Elsevier Inc. |
| Keywords: | genetics; dna replication; chromosome; metabolism; chromatin; dna helicases; dna helicase; chromosomes; replication termination; rdna; nanopore sequencing; replicon-seq; replisome pausing; rrm3; sister replisome |
| Journal Title: | Molecular Cell |
| Volume: | 82 |
| Issue: | 7 |
| ISSN: | 1097-2765 |
| Publisher: | Cell Press |
| Date Published: | 2022-04-07 |
| Start Page: | 1372 |
| End Page: | 1382.e4 |
| Language: | English |
| DOI: | 10.1016/j.molcel.2022.02.010 |
| PUBMED: | 35240057 |
| PROVIDER: | scopus |
| PMCID: | PMC8995386 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 2 May 2022 -- Source: Scopus |
Altmetric
Citation Impact
BMJ Impact Analytics
MSK Authors
-
16Whitehouse -
2Claussin -
1Vazquez
Related MSK Work