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The fork protection complex recruits FACT to reorganize nucleosomes during replication

Results of the Kurat Lab's collaboration with the Duderstadt lab and colleagues in Nucleic Acids Research


Barbara Safaric, Erika Chacin, Matthias J Scherr, Lional Rajappa, Christian Gebhardt, Christoph F Kurat, Thorben Cordes, Karl E Duderstadt (Jan 21, 2022) Nucleic Acids Research, gkac005.  https://doi.org/10.1093/nar/gkac005

Abstract cited from article:

Chromosome replication depends on efficient removal of nucleosomes by accessory factors to ensure rapid access to genomic information. Here, we show this process requires recruitment of the nucleosome reorganization activity of the histone chaperone FACT. Using single-molecule FRET, we demonstrate that reorganization of nucleosomal DNA by FACT requires coordinated engagement by the middle and C-terminal domains of Spt16 and Pob3 but does not require the N-terminus of Spt16. Using structure-guided pulldowns, we demonstrate instead that the N-terminal region is critical for recruitment by the fork protection complex subunit Tof1. Using in vitro chromatin replication assays, we confirm the importance of these interactions for robust replication. Our findings support a mechanism in which nucleosomes are removed through the coordinated engagement of multiple FACT domains positioned at the replication fork by the fork protection complex.