Martin BJE, Chruscicki AT, Howe LJ

The FACT (FAcilitates Chromatin Transactions) complex is enriched on highly expressed genes, where it facilitates transcription while maintaining chromatin structure. How it is targeted to these regions is unknown. In vitro, FACT binds destabilized nucleosomes, supporting the hypothesis that FACT is targeted to transcribed chromatin through recognition of RNA polymerase-disrupted ...

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nucleosomes. In this study, we used high resolution analysis of FACT occupancy in Saccharomyces cerevisiae to test this hypothesis. We demonstrate that FACT interacts with destabilized nucleosomes in vivo and its interaction with chromatin is dependent on transcription by any of the three RNA polymerases. Deep sequencing of micrococcal nuclease-resistant fragments shows that FACT-bound nucleosomes exhibit differences in micrococcal nuclease sensitivity compared to bulk chromatin, consistent with a modified nucleosome structure being the preferred ligand for this complex. Interestingly, a subset of FACT-bound nucleosomes may be "overlapping di-nucleosomes", in which one histone octamer invades the ~147 bp territory normally occupied by the adjacent nucleosome. While the presence of altered nucleosomes associated with FACT can also be explained by the known ability of this complex to reorganize nucleosome structure, transcription inhibition alleviates this effect indicating that it is not due to FACT interaction alone. Collectively these results suggest that FACT is targeted to transcribed genes through preferential interaction with RNA polymerase-disrupted nucleosomes.

Date: Sep. 20, 2018

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