Molecular mechanisms controlling Rif1 association to chromatin in mouse embryonic stem cells

Supervisors: Sara Buonomo, Andrew Goryachev

Project description:

DNA replication is a key event for the transmission of genetic information. The eukaryotic genome does not replicate all at once, but it follows a temporal program that reproducibly ensures replication of different parts of the genome at different stages of S-phase. This is called the DNA replication-timing program. We have discovered Rif1 as the major player in the control of replication timing as well as a key link with chromatin three-dimensional organisation in the nucleus. However, we do not know how Rif1 dynamic association to the chromatin is regulated. The aim of this project is to measure the dynamics of Rif1 association to chromatin in vivo in mouse embryonic stem cells taking advantage of a GFP-knock-in allele generated in the lab.  We will perform FRAP and FLIP experiments to monitor the changes to Rif1’s chromatin-association dynamics during cell cycle and model it.  

In parallel we will undertake a biochemical approach to characterize the chromatin features that drive Rif1 binding to the late replicating genomic regions and a genetic approach to explore the role of the characterised Rif1 DNA binding domain.

If you are interested in cell cycle, DNA replication, nuclear organization and want to work with mammalian primary and stem cells join our group. We are looking for enthusiastic, flexible and hard working candidates driven by curiosity and passion for science. Basic knowledge of cell/molecular biology and ability to work in English, in an international environment are required.

References:

Sukackaite et al., Mouse Rif2 is a regulatory subunit of protein phosphatase 1 (PP1), Sci Rep. 2017 May 18;7(1):2119

Foti R. et al., Nuclear architecture organized by Rif1 underpins the Replication-timing program,. Mol Cell. 2016 Jan 21;61(2):260-73.

Cornacchia D et al., Mouse Rif1 is a key regulator of the DNA replication-timing program in mammalian cells, EMBO J. 2012 Sep 12;31(18):3678-90.

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