Polyubiquitinated PCNA Recruits the ZRANB3 Translocase to Maintain Genomic Integrity after Replication Stress

Department of Genetics, Harvard University Medical School, Boston, MA 02115, USA.
Molecular cell (Impact Factor: 14.02). 06/2012; 47(3):396-409. DOI: 10.1016/j.molcel.2012.05.024
Source: PubMed


Completion of DNA replication after replication stress depends on PCNA, which undergoes monoubiquitination to stimulate direct bypass of DNA lesions by specialized DNA polymerases or is polyubiquitinated to promote recombination-dependent DNA synthesis across DNA lesions by template switching mechanisms. Here we report that the ZRANB3 translocase, a SNF2 family member related to the SIOD disorder SMARCAL1 protein, is recruited by polyubiquitinated PCNA to promote fork restart following replication arrest. ZRANB3 depletion in mammalian cells results in an increased frequency of sister chromatid exchange and DNA damage sensitivity after treatment with agents that cause replication stress. Using in vitro biochemical assays, we show that recombinant ZRANB3 remodels DNA structures mimicking stalled replication forks and disassembles recombination intermediates. We therefore propose that ZRANB3 maintains genomic stability at stalled or collapsed replication forks by facilitating fork restart and limiting inappropriate recombination that could occur during template switching events.

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Available from: Lior Izhar, May 04, 2015
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    • "In response to replication blocks, PCNA undergoes monoubiquitination that recruits translesion polymerases and stimulates direct replication bypass. Additionally , PCNA can become polyubiquitinated with K63-linked chains to promote recombination-dependent DNA synthesis across DNA lesions by template-switching mechanisms and recruits ZRANB3 to sites of replication stress (Ciccia et al., 2012). FANCI and FANCD2 respond to DNA interstrand crosslinks that arrest replication forks and are recruited to damaged DNA where they are ubiquitinated and promote repair. "
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    • "It will also be important to identify specific enzymatic activities required to assist RAD51 in driving replication fork reversal in vivo (Fig. 8; Neelsen and Lopes, 2015), presumably included in the list of factors showing fork remodeling activity in vitro (Kanagaraj et al., 2006; Machwe et al., 2006; Ralf et al., 2006; Gari et al., 2008; Blastyák et al., 2010; Bugreev et al., 2011; Bétous et al., 2012, 2013; Ciccia et al., 2012; Burkovics et al., 2014). Conversely, in light of our data, it will be important to extend the limited information on how the addition of RAD51 and RPA in the reactions may impact the biochemical properties of these fork remodeling proteins (Kanagaraj et al., 2006; Bugreev et al., 2011; Bétous et al., 2013; Burkovics et al., 2014). "
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    • "More than 400 proteins may interact with PCNA using two binding motifs, PCNA-interacting peptide (PIP)‐box [9] and AlkB homologue 2 PCNA-interacting motif (APIM) [10]. We and others have demonstrated that functional interaction with PCNA is mediated through APIM in several DNA repair proteins important for repair of ICLs [10] [11] [12] [13] [14]. Hence, several lines of evidence suggest that PCNA interactions through the APIM sequence are vital during cellular stress. "
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