Histone H3 1 serine-57 is a CHK1 substrate whose phosphorylation controls DNA repair Nikolaos Parisis, Pablo D Dans, Muhammad Jbara, Balveer Singh, Diane Schausi-Tiffoche, Diego Molina-Serrano, Isabelle Brun-Heath, Denisa Hendrychová, Suman Kumar Maity, Diana Buitrago, Rafael Lema, Thiziri Nait Achour, Simona Giunta, Michael Girardot, Nicolas Talarek, Valérie Rofidal, Katerina Danezi, Damien Coudreuse, Marie-Noëlle Prioleau, Robert Feil, Modesto Orozco, Ashraf Brik, Pei-Yun Jenny Wu, Liliana Krasinska, Daniel Fisher Nature Communications 2023, Aug 22;14(1):5104. doi: 10.1038/s41467-023-40843-4

 

2023-08-15

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Histone post-translational modifications promote a chromatin environment that controls transcription, DNA replication and repair, but surprisingly few phosphorylations have been documented. We report the discovery of histone H3 serine-57 phosphorylation (H3S57ph) and show that it is implicated in different DNA repair pathways from fungi to vertebrates. We identified CHK1 as a major human H3S57 kinase, and disrupting or constitutively mimicking H3S57ph had opposing effects on rate of recovery from replication stress, 53BP1 chromatin binding, and dependency on RAD52. In fission yeast, mutation of all H3 alleles to S57A abrogated DNA repair by both non-homologous end-joining and homologous recombination, while cells with phospho-mimicking S57D alleles were partly compromised for both repair pathways, presented aberrant Rad52 foci and were strongly sensitised to replication stress. Mechanistically, H3S57ph loosens DNA-histone contacts, increasing nucleosome mobility, and interacts with H3K56. Our results suggest that dynamic phosphorylation of H3S57 is required for DNA repair and recovery from replication stress, opening avenues for investigating the role of this modification in other DNA-related processes.


Long-term evolution of proliferating cells using the eVOLVER platform Daniel García-Ruano, Akanksha Jain, Zachary J. Heins, Brandon G. Wong, Ezira Yimer Wolle, Ahmad S. Khalil and Damien Coudreuse Open Biology 2023 Jul;13(7):230118. doi: 10.1098/rsob.230118

 

2023-06-28

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Experimental evolution using fast-growing unicellular organisms is a unique strategy for deciphering the principles and mechanisms underlying evolutionary processes as well as the architecture and wiring of basic biological functions.Over the past decade, this approach has benefited from the development of powerful systems for the continuous control of the growth of independently evolving cultures. While the first devices compatible with multiplexed experimental evolution remained challenging to implement and required constant user intervention, the recently developed eVOLVER framework represents a fully automated closed-loop system for laboratory evolution assays. However, it remained difficult to maintain and compare parallel evolving cultures in tightly controlled environments over long periods of time using eVOLVER. Furthermore, a number of tools were lacking to cope with the various issues that inevitably occur when conducting such long-term assays. Here we present a significant upgrade of the eVOLVER framework, providing major modifications of the experimental methodology, hardware and software as well as a new stand-alone protocol. Altogether, these adaptations and improvements make the eVOLVER a versatile and unparalleled set-up for long-term experimental evolution.


Fluorescence exclusion - a rapid, accurate and powerful method for measuring yeast cell volume Daniel García-Ruano, Larisa Venkova, Akanksha Jain, Joseph C Ryan, Vasanthakrishnan Radhakrishnan Balasubramaniam, Matthieu Piel, Damien Coudreuse J Cell Sci. 2022 Jul 1;135(13):jcs259392. doi: 10.1242/jcs.259392. Epub 2022 Jun 30.

 

2022-07-01

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Cells exist in an astonishing range of volumes across and within species. However, our understanding of cell size control remains limited, owing in large part to the challenges associated with accurate determination of cell volume. Much of our comprehension of size regulation derives from yeast models, but even for these morphologically stereotypical cells, assessment of cell volume has mostly relied on proxies and extrapolations from two-dimensional measurements. Recently, the fluorescence exclusion method (FXm) was developed to evaluate the size of mammalian cells, but whether it could be applied to smaller cells remained unknown. Using specifically designed microfluidic chips and an improved data analysis pipeline, we show here that FXm reliably detects subtle differences in the volume of fission yeast cells, even for those with altered shapes. Moreover, it allows for the monitoring of dynamic volume changes at the single-cell level with high time resolution. Collectively, our work highlights how the coupling of FXm with yeast genetics will bring new insights into the complex biology of cell growth.