The Histone Chaperone Facilitates Chromatin Transcription (FACT) Protein Maintains Normal Replication Fork Rates

Ordered nucleosome disassembly and reassembly are required for eukaryotic DNA replication. The facilitates chromatin transcription (FACT) complex, a histone chaperone comprising Spt16 and SSRP1, is involved in DNA replication as well as transcription. FACT associates with the MCM helicase, which is...

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Published in:The Journal of biological chemistry 2011-09, Vol.286 (35), p.30504-30512
Main Authors: Abe, Takuya, Sugimura, Kazuto, Hosono, Yoshifumi, Takami, Yasunari, Akita, Motomu, Yoshimura, Akari, Tada, Shusuke, Nakayama, Tatsuo, Murofushi, Hiromu, Okumura, Katsuzumi, Takeda, Shunichi, Horikoshi, Masami, Seki, Masayuki, Enomoto, Takemi
Format: Article
Language:English
Subjects:
Animals
Cell Cycle
Chickens
Chromatin
Chromatin - chemistry
DNA and Chromosomes
DNA Replication
DNA-Binding Proteins - metabolism
Epigenesis, Genetic
Epigenetics
Flow Cytometry - methods
High Mobility Group Proteins - metabolism
Histone Chaperone
Histone Chaperones - chemistry
Histones - chemistry
Humans
Molecular Chaperones - metabolism
Nucleosome
Nucleosomes - metabolism
S Phase
Transcription, Genetic
Transcriptional Elongation Factors - metabolism
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Summary:Ordered nucleosome disassembly and reassembly are required for eukaryotic DNA replication. The facilitates chromatin transcription (FACT) complex, a histone chaperone comprising Spt16 and SSRP1, is involved in DNA replication as well as transcription. FACT associates with the MCM helicase, which is involved in DNA replication initiation and elongation. Although the FACT-MCM complex is reported to regulate DNA replication initiation, its functional role in DNA replication elongation remains elusive. To elucidate the functional role of FACT in replication fork progression during DNA elongation in the cells, we generated and analyzed conditional SSRP1 gene knock-out chicken (Gallus gallus) DT40 cells. SSRP1-depleted cells ceased to grow and exhibited a delay in S-phase cell cycle progression, although SSRP1 depletion did not affect the level of chromatin-bound DNA polymerase α or nucleosome reassembly on daughter strands. The tracking length of newly synthesized DNA, but not origin firing, was reduced in SSRP1-depleted cells, suggesting that the S-phase cell cycle delay is mainly due to the inhibition of replication fork progression rather than to defects in the initiation of DNA replication in these cells. We discuss the mechanisms of how FACT promotes replication fork progression in the cells.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M111.264721