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Biochimica et Biophysica Acta (BBA) - Molecular Cell Research
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Biochimica et Biophysica Acta (BBA) - Molecular Cell Research
Article . 2011 . Peer-reviewed
License: Elsevier Non-Commercial
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Biphasic chromatin binding of histone chaperone FACT during eukaryotic chromatin DNA replication

Authors: Kundu, Lena R.; Seki, Masayuki; Watanabe, Nanae; Murofushi, Hiromu; Furukohri, Asako; Waga, Shou; Score, Alan J.; +4 Authors

Biphasic chromatin binding of histone chaperone FACT during eukaryotic chromatin DNA replication

Abstract

The facilitates chromatin transcription (FACT) complex affects nuclear DNA transactions in a chromatin context. Though the involvement of FACT in eukaryotic DNA replication has been revealed, a clear understanding of its biochemical behavior during DNA replication still remains elusive. Here, we analyzed the chromatin-binding dynamics of FACT using Xenopus egg extract cell-free system. We found that FACT has at least two distinct chromatin-binding phases: (1) a rapid chromatin-binding phase at the onset of DNA replication that did not involve origin licensing and (2) a second phase of chromatin binding that initiated after origin licensing. Intriguingly, early-binding FACT dissociated from chromatin when DNA replication was blocked by the addition of Cdc6 in the licensed state before origin firing. Cdc6-induced removal of FACT was blocked by the inhibition of origin licensing with geminin, but not by suppressing the activity of DNA polymerases, CDK, or Cdc7. Furthermore, chromatin transfer experiments revealed that impairing the later binding of FACT severely compromises DNA replication activity. Taken together, we propose that even though FACT has rapid chromatin-binding activity, the binding pattern of FACT on chromatin changes after origin licensing, which may contribute to the establishment of its functional link to the DNA replication machinery.

Country
United Kingdom
Keywords

DNA Replication, Male, 570, Time Factors, Origin licensing, Immunoblotting, FACT, 610, DUF, Cell Cycle Proteins, DNA replication, SSRP1, CELL-FREE-EXTRACTS, INITIATION, Animals, Humans, Histone Chaperones, Molecular Biology, Glutathione Transferase, FUNCTIONAL COOPERATION, Spt16, High Mobility Group Proteins, Nuclear Proteins, IN-VITRO, Cell Biology, ORIGIN RECOGNITION COMPLEX, Spermatozoa, POLYMERASE ALPHA, Chromatin, DNA-Binding Proteins, Kinetics, S-PHASE, XENOPUS, Eukaryotic Cells, LICENSING SYSTEM, Oocytes, MCM-HELICASE, Female, Transcriptional Elongation Factors, Protein Binding

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citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
9
Average
Average
Average
hybrid