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Mol Cell Biol. 2016 Mar 31;36(8):1287-96. doi: 10.1128/MCB.00835-15. Print 2016 Apr.

Histone Chaperone Nap1 Is a Major Regulator of Histone H2A-H2B Dynamics at the Inducible GAL Locus.

Author information

1
Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, Colorado, USA.
2
Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, Colorado, USA Howard Hughes Medical Institute, Colorado State University, Fort Collins, Colorado, USA.
3
Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, Colorado, USA Howard Hughes Medical Institute, Colorado State University, Fort Collins, Colorado, USA Institute for Genome Architecture and Function, Colorado State University, Fort Collins, Colorado, USA.
4
Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, Colorado, USA Institute for Genome Architecture and Function, Colorado State University, Fort Collins, Colorado, USA Laurie.Stargell@colostate.edu.

Abstract

Histone chaperones, like nucleosome assembly protein 1 (Nap1), play a critical role in the maintenance of chromatin architecture. Here, we use the GAL locus in Saccharomyces cerevisiae to investigate the influence of Nap1 on chromatin structure and histone dynamics during distinct transcriptional states. When the GAL locus is not expressed, cells lacking Nap1 show an accumulation of histone H2A-H2B but not histone H3-H4 at this locus. Excess H2A-H2B interacts with the linker DNA between nucleosomes, and the interaction is independent of the inherent DNA-binding affinity of H2A-H2B for these particular sequences as measured in vitro When the GAL locus is transcribed, excess H2A-H2B is reversed, and levels of all chromatin-bound histones are depleted in cells lacking Nap1. We developed an in vivo system to measure histone exchange at the GAL locus and observed considerable variability in the rate of exchange across the locus in wild-type cells. We recapitulate this variability with in vitro nucleosome reconstitutions, which suggests a contribution of DNA sequence to histone dynamics. We also find that Nap1 is required for transcription-dependent H2A-H2B exchange. Altogether, these results indicate that Nap1 is essential for maintaining proper chromatin composition and modulating the exchange of H2A-H2B in vivo.

PMID:
26884462
PMCID:
PMC4836277
DOI:
10.1128/MCB.00835-15
[Indexed for MEDLINE]
Free PMC Article

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