Dbf4 recruitment by forkhead transcription factors defines an upstream rate-limiting step in determining origin firing timing (original) (raw)

1. Armelle Lengronne2,5,
2. Di Shi1,5,
3. Romain Forey2,
4. Magdalena Skrzypczak3,
5. Krzysztof Ginalski3,
6. Changhui Yan4,
7. Xiaoke Wang1,
8. Qinhong Cao1,
9. Philippe Pasero2 and
10. Huiqiang Lou1
11. 1State Key Laboratory of Agro-Biotechnology, Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Biological Sciences, China Agricultural University, Beijing 100193, China;
12. 2Institute of Human Genetics, Centre National de la Recherche Scientifique, University of Montpellier, Equipe Labellisée Ligue Contre le Cancer, F-34396 Montpellier Cedex 5, France;
13. 3Laboratory of Bioinformatics and Systems Biology, Centre of New Technologies, University of Warsaw, 02-089 Warsaw, Poland;
14. 4Department of Computer Science, North Dakota State University, Fargo, North Dakota 58108, USA
15. Corresponding authors: lou{at}cau.edu.cn, philippe.pasero{at}igh.cnrs.fr
16. ↵5 These authors contributed equally to this work.

Abstract

Initiation of eukaryotic chromosome replication follows a spatiotemporal program. The current model suggests that replication origins compete for a limited pool of initiation factors. However, it remains to be answered how these limiting factors are preferentially recruited to early origins. Here, we report that Dbf4 is enriched at early origins through its interaction with forkhead transcription factors Fkh1 and Fkh2. This interaction is mediated by the Dbf4 C terminus and was successfully reconstituted in vitro. An interaction-defective mutant, dbf4ΔC, phenocopies fkh alleles in terms of origin firing. Remarkably, genome-wide replication profiles reveal that the direct fusion of the DNA-binding domain (DBD) of Fkh1 to Dbf4 restores the Fkh-dependent origin firing but interferes specifically with the pericentromeric origin activation. Furthermore, Dbf4 interacts directly with Sld3 and promotes the recruitment of downstream limiting factors. These data suggest that Fkh1 targets Dbf4 to a subset of noncentromeric origins to promote early replication in a manner that is reminiscent of the recruitment of Dbf4 to pericentromeric origins by Ctf19.

• DNA replication timing
• limiting factors
• Dbf4
• forkhead transcription factor

Footnotes

• Supplemental material is available for this article.

• Article published online ahead of print. Article and publication date are online at http://www.genesdev.org/cgi/doi/10.1101/gad.306571.117.

• Received August 28, 2017.

• Accepted November 30, 2017.

• © 2018 Fang et al.; Published by Cold Spring Harbor Laboratory Press

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