Identification of an ubinuclein 1 region required for stability and function of the human HIRA/UBN1/CABIN1/ASF1a histone H3.3 chaperone complex

Yong Tang, Aastha Puri, M. Daniel Ricketts, Taranjit Singh Rai, Jason Hoffmann, Elise Hoi, Peter D. Adams, David C. Schultz, Ronen Marmorstein

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

The mammalian HIRA/UBN1/CABIN1/ASF1a (HUCA) histone chaperone complex deposits the histone H3 variant H3.3 into chromatin and is linked to gene activation, repression, and chromatin assembly in diverse cell contexts. We recently reported that a short N-terminal fragment of UBN1 containing amino acids 1-175 is necessary and sufficient for interaction with the WD repeats of HIRA and attributed this interaction to a region from residues 120-175 that is highly conserved with the yeast ortholog Hpc2 and so termed the HRD for Hpc2-related domain. In this report, through a more comprehensive and refined biochemical and mutational analysis, we identify a smaller and more moderately conserved region within residues 41-77 of UBN1, which we term the NHRD, that is essential for interaction with the HIRA WD repeats; we further demonstrate that the HRD is dispensable for this interaction. We employ analytical ultracentrifugation studies to demonstrate that the NHRD of UBN1 and the WD repeats of HIRA form a tight 1:1 complex with a dissociation constant in the nanomolar range. Mutagenesis experiments identify several key residues in the NHRD that are required for interaction with the HIRA WD repeat domain, stability of the HUCA complex in vitro and in vivo, and changes in chromatin organization in primary human cells. Together, these studies implicate the NHRD domain of UBN1 as being an essential region for HIRA interaction and chromatin organization by the HUCA complex.

LanguageEnglish
Pages2366-2377
Number of pages12
JournalBiochemistry
Volume51
Issue number12
DOIs
Publication statusPublished - 27 Mar 2012

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Histone Chaperones
Histones
Chromatin
Mutagenesis
Chromatin Assembly and Disassembly
Ultracentrifugation
Yeast
Transcriptional Activation
Deposits
Genes
Yeasts
Chemical activation
Cells
Amino Acids
WD40 Repeats
Experiments

Cite this

Tang, Yong ; Puri, Aastha ; Ricketts, M. Daniel ; Rai, Taranjit Singh ; Hoffmann, Jason ; Hoi, Elise ; Adams, Peter D. ; Schultz, David C. ; Marmorstein, Ronen. / Identification of an ubinuclein 1 region required for stability and function of the human HIRA/UBN1/CABIN1/ASF1a histone H3.3 chaperone complex. In: Biochemistry. 2012 ; Vol. 51, No. 12. pp. 2366-2377.
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abstract = "The mammalian HIRA/UBN1/CABIN1/ASF1a (HUCA) histone chaperone complex deposits the histone H3 variant H3.3 into chromatin and is linked to gene activation, repression, and chromatin assembly in diverse cell contexts. We recently reported that a short N-terminal fragment of UBN1 containing amino acids 1-175 is necessary and sufficient for interaction with the WD repeats of HIRA and attributed this interaction to a region from residues 120-175 that is highly conserved with the yeast ortholog Hpc2 and so termed the HRD for Hpc2-related domain. In this report, through a more comprehensive and refined biochemical and mutational analysis, we identify a smaller and more moderately conserved region within residues 41-77 of UBN1, which we term the NHRD, that is essential for interaction with the HIRA WD repeats; we further demonstrate that the HRD is dispensable for this interaction. We employ analytical ultracentrifugation studies to demonstrate that the NHRD of UBN1 and the WD repeats of HIRA form a tight 1:1 complex with a dissociation constant in the nanomolar range. Mutagenesis experiments identify several key residues in the NHRD that are required for interaction with the HIRA WD repeat domain, stability of the HUCA complex in vitro and in vivo, and changes in chromatin organization in primary human cells. Together, these studies implicate the NHRD domain of UBN1 as being an essential region for HIRA interaction and chromatin organization by the HUCA complex.",
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Tang, Y, Puri, A, Ricketts, MD, Rai, TS, Hoffmann, J, Hoi, E, Adams, PD, Schultz, DC & Marmorstein, R 2012, 'Identification of an ubinuclein 1 region required for stability and function of the human HIRA/UBN1/CABIN1/ASF1a histone H3.3 chaperone complex', Biochemistry, vol. 51, no. 12, pp. 2366-2377. https://doi.org/10.1021/bi300050b

Identification of an ubinuclein 1 region required for stability and function of the human HIRA/UBN1/CABIN1/ASF1a histone H3.3 chaperone complex. / Tang, Yong; Puri, Aastha; Ricketts, M. Daniel; Rai, Taranjit Singh; Hoffmann, Jason; Hoi, Elise; Adams, Peter D.; Schultz, David C.; Marmorstein, Ronen.

In: Biochemistry, Vol. 51, No. 12, 27.03.2012, p. 2366-2377.

Research output: Contribution to journalArticle

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T1 - Identification of an ubinuclein 1 region required for stability and function of the human HIRA/UBN1/CABIN1/ASF1a histone H3.3 chaperone complex

AU - Tang, Yong

AU - Puri, Aastha

AU - Ricketts, M. Daniel

AU - Rai, Taranjit Singh

AU - Hoffmann, Jason

AU - Hoi, Elise

AU - Adams, Peter D.

AU - Schultz, David C.

AU - Marmorstein, Ronen

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N2 - The mammalian HIRA/UBN1/CABIN1/ASF1a (HUCA) histone chaperone complex deposits the histone H3 variant H3.3 into chromatin and is linked to gene activation, repression, and chromatin assembly in diverse cell contexts. We recently reported that a short N-terminal fragment of UBN1 containing amino acids 1-175 is necessary and sufficient for interaction with the WD repeats of HIRA and attributed this interaction to a region from residues 120-175 that is highly conserved with the yeast ortholog Hpc2 and so termed the HRD for Hpc2-related domain. In this report, through a more comprehensive and refined biochemical and mutational analysis, we identify a smaller and more moderately conserved region within residues 41-77 of UBN1, which we term the NHRD, that is essential for interaction with the HIRA WD repeats; we further demonstrate that the HRD is dispensable for this interaction. We employ analytical ultracentrifugation studies to demonstrate that the NHRD of UBN1 and the WD repeats of HIRA form a tight 1:1 complex with a dissociation constant in the nanomolar range. Mutagenesis experiments identify several key residues in the NHRD that are required for interaction with the HIRA WD repeat domain, stability of the HUCA complex in vitro and in vivo, and changes in chromatin organization in primary human cells. Together, these studies implicate the NHRD domain of UBN1 as being an essential region for HIRA interaction and chromatin organization by the HUCA complex.

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