A non-transcriptional function of Yap regulates the DNA replication program
- Paris-Saclay Institute of Neuroscience, France
- CNRS, CEA, University Paris Sud, France
Abstract
In multicellular eukaryotic organisms, the initiation of DNA replication occurs asynchronously throughout S-phase according to a regulated replication timing program. Here, using Xenopus egg extracts, we showed that Yap (Yes-associated protein 1), a downstream effector of the Hippo signalling pathway, is required for the control of DNA replication dynamics. We found that Yap is recruited to chromatin at the start of DNA replication and identified Rif1, a major regulator of the DNA replication timing program, as a novel Yap binding protein. Furthermore, we show that either Yap or Rif1 depletion accelerates DNA replication dynamics by increasing the number of activated replication origins. In Xenopus embryos, using a Trim-Away approach during cleavage stages devoid of transcription, we found that either Yap or Rif1 depletion triggers an acceleration of cell divisions, suggesting a shorter S-phase by alterations of the replication program. Finally, our data show that Rif1 knockdown leads to defects in the partitioning of early versus late replication foci in retinal stem cells, as we previously showed for Yap. Altogether, our findings unveil a non-transcriptional role for Yap in regulating replication dynamics. We propose that Yap and Rif1 function as breaks to control the DNA replication program in early embryos and post-embryonic stem cells.
Data availability
Source data files have been provided for all Western blots and graphs shown on the figures.We have submitted our dataset "Identification of Yap-interacting proteins in Xenopus egg extracts by co-immunoprecipitation coupled to LC/MS/MS" to ProteomeXchange via the PRIDE database (Project accession: PXD029345; Project DOI: 10.6019/PXD029345).
Article and author information
Author details
Kathrin Marheineke
Funding
Association pour la Recherche sur le Cancer (NA)
- Muriel Perron
- Odile Bronchain
Retina France (NA)
- Muriel Perron
Fondation Valentin Haüy (NA)
- Muriel Perron
UNADEV (NA)
- Muriel Perron
Conacyt (439641)
- Rodrigo Meléndez García
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: All animal experiments have been carried out in accordance with the European Community Council Directive of 22 September 2010 (2010/63/EEC). All animal care and experimentation were conducted in accordance with institutional guidelines, under the institutional license C 91-471-102. The study protocols were approved by the institutional animal care committee CEEA #59 and received an authorization by the Direction Départementale de la Protection des Populations under the reference APAFIS#998-2015062510022908v2 for Xenopus experiments.
Copyright
© 2022, Meléndez García et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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A non-transcriptional function of Yap regulates the DNA replication program
eLife 11:e75741.
https://doi.org/10.7554/eLife.75741
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