Processing of DNA Polymerase-Blocking Lesions during Genome Replication Is Spatially and Temporally Segregated from Replication Forks
Tracing DNA repair factors by fluorescence microscopy provides valuable information about how DNA damage processing is orchestrated within cells. Most repair pathways involve single-stranded DNA (ssDNA), making replication protein A (RPA) a hallmark of DNA damage and replication stress. RPA foci eme...
| Autores: | , , , , |
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| Tipo de recurso: | artículo |
| Estado: | Versión aceptada para publicación |
| Fecha de publicación: | 2020 |
| País: | España |
| Institución: | Universidad de Sevilla (US) |
| Repositorio: | idUS. Depósito de Investigación de la Universidad de Sevilla |
| OAI Identifier: | oai:idus.us.es:11441/166496 |
| Acceso en línea: | https://hdl.handle.net/11441/166496 https://doi.org/10.1016/j.molcel.2019.09.015 |
| Access Level: | acceso abierto |
| Palabra clave: | Daughter-strand gaps DNA damage bypass DNA replication DNA replication stress Homologous recombination PCNA RAD6 pathway RPA Translesion synthesis Ubiquitin |
| Sumario: | Tracing DNA repair factors by fluorescence microscopy provides valuable information about how DNA damage processing is orchestrated within cells. Most repair pathways involve single-stranded DNA (ssDNA), making replication protein A (RPA) a hallmark of DNA damage and replication stress. RPA foci emerging during S phase in response to tolerable loads of polymerase-blocking lesions are generally thought to indicate stalled replication intermediates. We now report that in budding yeast they predominantly form far away from sites of ongoing replication, and they do not overlap with any of the repair centers associated with collapsed replication forks or double-strand breaks. Instead, they represent sites of postreplicative DNA damage bypass involving translesion synthesis and homologous recombination. We propose that most RPA and recombination foci induced by polymerase-blocking lesions in the replication template are clusters of repair tracts arising from replication centers by polymerase re-priming and subsequent expansion of daughter-strand gaps over the course of S phase. © 2019 Elsevier Inc. Wong et al. use the single-stranded DNA-binding RPA complex as a reporter of how replication of damaged DNA is orchestrated in the budding yeast nucleus. In response to polymerase-blocking lesions, local clusters of single-stranded DNA originate in the wake of replication forks, to be resolved by gap-filling in postreplicative repair territories (PORTs). |
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