Cac1 WHD and PIP domains have distinct roles in replisome progression and genomic stability
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ORIGINAL ARTICLE
Cac1 WHD and PIP domains have distinct roles in replisome progression and genomic stability Ioannis Tsirkas1 · Daniel Dovrat1 · Yang Lei2 · Angeliki Kalyva1 · Diana Lotysh1 · Qing Li2 · Amir Aharoni1 Received: 1 July 2020 / Revised: 17 September 2020 / Accepted: 21 September 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Replication-coupled (RC) nucleosome assembly is an essential process in eukaryotic cells to maintain chromatin structure during DNA replication. The deposition of newly-synthesized H3/H4 histones during DNA replication is facilitated by specialized histone chaperones. CAF-1 is an important histone chaperone complex and its main subunit, Cac1p, contains a PIP and WHD domain for interaction with PCNA and the DNA, respectively. While Cac1p subunit was extensively studied in different systems much less is known regarding the importance of the PIP and WHD domains in replication fork progression and genome stability. By exploiting a time-lapse microscopy system for monitoring DNA replication in individual live cells, we examined how mutations in these Cac1p domains affect replication fork progression and post-replication characteristics. Our experiments revealed that mutations in the Cac1p WHD domain, which abolished the CAF-1–DNA interaction, slows down replication fork progression. In contrast, mutations in Cac1p PIP domain, abolishing Cac1p–PCNA interaction, lead to extended late-S/Anaphase duration, elevated number of RPA foci and increased spontaneous mutation rate. Our research shows that Cac1p WHD and PIP domains have distinct roles in high replisome progression and maintaining genome stability during cell cycle progression. Keywords DNA replication · Histone chaperones · Cac1
Introduction DNA replication in eukaryotic cells is a complex process that requires the accurate copying of the DNA itself and the formation of a precise chromatin structure (Alabert and Groth 2012; Papamichos-Chronakis and Peterson 2012). The basic unit of the chromatin is the nucleosome, composed of ~ 146 base pairs of DNA wrapped around an Communicated by M. Kupiec. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00294-020-01113-8) contains supplementary material, which is available to authorized users. * Amir Aharoni [email protected] 1
Department of Life Sciences and the National Institute for Biotechnology in the Negev, Ben-Gurion University of the Negev, 84105 Be’er Sheva, Israel
State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences and Peking‑Tsinghua Center for Life Sciences, Peking University, Beijing 100871, China
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octamer of histones. A nucleosome is composed of a core of (H3-H4)2 tetramer and two flanking H2A-H2B dimmers (Falbo and Shen 2006; Andrews and Luger 2011). During DNA replication, nucleosomes must be disassembled to allow replication fork progression and subsequently must be reassembled to establish the accurate chromatin state. Histone chaperones are essential for
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