Identification of Elg1 interaction partners and effects on post-replication chromatin re-formation

• Published in: PLoS genetics 2018-11, Vol.14 (11), p.e1007783-e1007783
• Main Authors: Gali, Vamsi K, Dickerson, David, Katou, Yuki, Fujiki, Katsunori, Shirahige, Katsuhiko, Owen-Hughes, Tom, Kubota, Takashi, Donaldson, Anne D
• Format: Article
• Language: English
• Subjects: Biology and Life Sciences; Cancer; Carrier Proteins - genetics; Carrier Proteins - metabolism; Cell cycle; Chromatin; Chromatin - genetics; Chromatin - metabolism; Deoxyribonucleic acid; DNA; DNA biosynthesis; DNA polymerase; DNA Replication; Epigenetics; Funding; Gene expression; Genes, Fungal; Genetic engineering; Genetic research; Genomes; Genomic Instability; Molecular Chaperones - genetics; Molecular Chaperones - metabolism; Mutation; Nuclease; Nucleases; Post-replication; Proliferating cell nuclear antigen; Proliferating Cell Nuclear Antigen - genetics; Proliferating Cell Nuclear Antigen - metabolism; Protein Binding; Proteomics; Replication factor C; Replication Protein C - genetics; Replication Protein C - metabolism; Research and Analysis Methods; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; Supervision; Transcription; Transcription (Genetics); Unloading; Yeast
• ISSN: 1553-7404; 1553-7390; 1553-7404
• DOI: 10.1371/journal.pgen.1007783

Elg1, the major subunit of a Replication Factor C-like complex, is critical to ensure genomic stability during DNA replication, and is implicated in controlling chromatin structure. We investigated the consequences of Elg1 loss for the dynamics of chromatin re-formation following DNA replication. Measurement of Okazaki fragment length and the micrococcal nuclease sensitivity of newly replicated DNA revealed a defect in nucleosome organization in the absence of Elg1. Using a proteomic approach to identify Elg1 binding partners, we discovered that Elg1 interacts with Rtt106, a histone chaperone implicated in replication-coupled nucleosome assembly that also regulates transcription. A central role for Elg1 is the unloading of PCNA from chromatin following DNA replication, so we examined the relative importance of Rtt106 and PCNA unloading for chromatin reassembly following DNA replication. We find that the major cause of the chromatin organization defects of an ELG1 mutant is PCNA retention on DNA following replication, with Rtt106-Elg1 interaction potentially playing a contributory role.