H3.Y discriminates between HIRA and DAXX chaperone complexes and reveals unexpected insights into human DAXX-H3.3-H4 binding and deposition requirements

TY - JOUR

T1 - H3.Y discriminates between HIRA and DAXX chaperone complexes and reveals unexpected insights into human DAXX-H3.3-H4 binding and deposition requirements

AU - Zink, Lisa-Maria

AU - Delbarre, Erwan

AU - Eberl, H Christian

AU - Keilhauer, Eva C

AU - Bönisch, Clemens

AU - Pünzeler, Sebastian

AU - Bartkuhn, Marek

AU - Collas, Philippe

AU - Mann, Matthias

AU - Hake, Sandra B

N1 - © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

PY - 2017/6/2

Y1 - 2017/6/2

N2 - Histone chaperones prevent promiscuous histone interactions before chromatin assembly. They guarantee faithful deposition of canonical histones and functionally specialized histone variants into chromatin in a spatial- and temporally-restricted manner. Here, we identify the binding partners of the primate-specific and H3.3-related histone variant H3.Y using several quantitative mass spectrometry approaches, and biochemical and cell biological assays. We find the HIRA, but not the DAXX/ATRX, complex to recognize H3.Y, explaining its presence in transcriptionally active euchromatic regions. Accordingly, H3.Y nucleosomes are enriched in the transcription-promoting FACT complex and depleted of repressive post-translational histone modifications. H3.Y mutational gain-of-function screens reveal an unexpected combinatorial amino acid sequence requirement for histone H3.3 interaction with DAXX but not HIRA, and for H3.3 recruitment to PML nuclear bodies. We demonstrate the importance and necessity of specific H3.3 core and C-terminal amino acids in discriminating between distinct chaperone complexes. Further, chromatin immunoprecipitation sequencing experiments reveal that in contrast to euchromatic HIRA-dependent deposition sites, human DAXX/ATRX-dependent regions of histone H3 variant incorporation are enriched in heterochromatic H3K9me3 and simple repeat sequences. These data demonstrate that H3.Y's unique amino acids allow a functional distinction between HIRA and DAXX binding and its consequent deposition into open chromatin.

AB - Histone chaperones prevent promiscuous histone interactions before chromatin assembly. They guarantee faithful deposition of canonical histones and functionally specialized histone variants into chromatin in a spatial- and temporally-restricted manner. Here, we identify the binding partners of the primate-specific and H3.3-related histone variant H3.Y using several quantitative mass spectrometry approaches, and biochemical and cell biological assays. We find the HIRA, but not the DAXX/ATRX, complex to recognize H3.Y, explaining its presence in transcriptionally active euchromatic regions. Accordingly, H3.Y nucleosomes are enriched in the transcription-promoting FACT complex and depleted of repressive post-translational histone modifications. H3.Y mutational gain-of-function screens reveal an unexpected combinatorial amino acid sequence requirement for histone H3.3 interaction with DAXX but not HIRA, and for H3.3 recruitment to PML nuclear bodies. We demonstrate the importance and necessity of specific H3.3 core and C-terminal amino acids in discriminating between distinct chaperone complexes. Further, chromatin immunoprecipitation sequencing experiments reveal that in contrast to euchromatic HIRA-dependent deposition sites, human DAXX/ATRX-dependent regions of histone H3 variant incorporation are enriched in heterochromatic H3K9me3 and simple repeat sequences. These data demonstrate that H3.Y's unique amino acids allow a functional distinction between HIRA and DAXX binding and its consequent deposition into open chromatin.

KW - Adaptor Proteins, Signal Transducing

KW - Amino Acid Sequence

KW - Animals

KW - Binding Sites

KW - Cell Cycle Proteins

KW - Cell Line, Transformed

KW - Chromatin

KW - DNA-Binding Proteins

KW - Epithelial Cells

KW - HeLa Cells

KW - High Mobility Group Proteins

KW - Histone Chaperones

KW - Histone Code

KW - Histones

KW - Humans

KW - Mesenchymal Stromal Cells

KW - Microsatellite Repeats

KW - Nuclear Proteins

KW - Nucleosomes

KW - Primary Cell Culture

KW - Protein Binding

KW - Sequence Alignment

KW - Sequence Homology, Amino Acid

KW - Transcription Factors

KW - Transcription, Genetic

KW - Transcriptional Elongation Factors

KW - Journal Article

U2 - 10.1093/nar/gkx131

DO - 10.1093/nar/gkx131

M3 - Journal article

C2 - 28334823

SN - 0305-1048

VL - 45

SP - 5691

EP - 5706

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 10

ER -