期刊
NUCLEIC ACIDS RESEARCH
卷 50, 期 3, 页码 1317-1334出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkac005
关键词
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资金
- H2020 European Research Council [804098]
- Max Planck Society
- Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) [213249687 -SFB 1064]
- European Research Council (ERC) [804098] Funding Source: European Research Council (ERC)
Chromosome replication relies on the recruitment of the nucleosome reorganization activity of the histone chaperone FACT, with coordination from the middle and C-terminal domains of Spt16 and Pob3. The N-terminal region plays a critical role in recruitment by the fork protection complex subunit Tof1. These interactions are essential for robust replication in vitro.
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.
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