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Structure of a human ASF1a–HIRA complex and insights into specificity of histone chaperone complex assembly

Nature Structural & Molecular Biology volume 13, pages 921–929 (2006)Cite this article

Abstract

Human HIRA, ASF1a, ASF1b and CAF-1 are evolutionally conserved histone chaperones that form multiple functionally distinct chromatin-assembly complexes, with roles linked to diverse nuclear process, such as DNA replication and formation of heterochromatin in senescent cells. We report the crystal structure of an ASF1a–HIRA heterodimer and a biochemical dissection of ASF1a's mutually exclusive interactions with HIRA and the p60 subunit of CAF-1. The HIRA B domain forms an antiparallel β-hairpin that binds perpendicular to the strands of the β-sandwich of ASF1a, via β-sheet, salt bridge and van der Waals contacts. The N- and C-terminal regions of ASF1a and ASF1b determine the different affinities of these two proteins for HIRA, by contacting regions outside the HIRA B domain. CAF-1 p60 also uses B domain–like motifs for binding to ASF1a, thereby competing with HIRA. Together, these studies begin to define the molecular determinants of assembly of functionally diverse macromolecular histone chaperone complexes.

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Figure 1: Sequence alignments of the B domains of HIRA and CAF-1 p60 homologs.
Figure 2: ASF1a binding to HIRA deletion constructs.
Figure 3: Overall structure of the ASF1a–HIRA complex and comparison with nascent yeast Asf1p and human ASF1a.
Figure 4: The ASF1a–HIRA interface.
Figure 5: Functional characterization of the ASF1a-HIRA interaction.
Figure 6: Functional characterization of ASF1a interaction with the HIRA B domain–like motifs of CAF-1 p60.
Figure 7: Model for binding of other HIRA regions and the histone H3.3 C-terminal helix to the HIRA B domain–ASF1a complex.

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Acknowledgements

We thank M. Allaire for assistance with data collection. This work was supported by US National Institutes of Health grants to R.M. and P.D.A., a Leukemia and Lymphoma Society Scholar Award to P.D.A. and a grant from the Commonwealth Universal Research Enhancement Program, Pennsylvania Department of Health, awarded to the Wistar Institute. Part of this research was conducted on beamline X6A at the National Synchrotron Light Source at Brookhaven National Laboratory, which is supported by the US Department of Energy under contract no. DE-AC02-98CH10886. Beamline X6A is funded by the US National Institutes of Health, National Institute of General Medical Sciences, under agreement Y1 GM-0080-03.

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Author notes

  1. Yong Tang and Maxim V Poustovoitov: These authors contributed equally to this work.

Authors and Affiliations

  1. The Wistar Institute, Philadelphia, 19104, Pennsylvania, USA

    Yong Tang, Kehao Zhao & Ronen Marmorstein

  2. The Fox Chase Cancer Center, Philadelphia, 19111, Pennsylvania, USA

    Maxim V Poustovoitov, Megan Garfinkel, Adrian Canutescu, Roland Dunbrack & Peter D Adams

  3. Russian State Medical University, Moscow, 117 869, Russia

    Maxim V Poustovoitov

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Contributions

Y.T. and M.V.P. designed and carried out the reported experiments and prepared manuscript figures and text; K.Z., M.G., A.C. and R.D. carried out preliminary studies that led to the reported experiments; P.D.A and R.M. designed and supervised experiments and prepared manuscript text. All authors read and approved the submitted manuscript.

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Correspondence to Peter D Adams or Ronen Marmorstein.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Sequence alignment of ASF1 homologs. (PDF 2466 kb)

Supplementary Fig. 2

ASF1a binding to HIRA deletion constructs. (PDF 558 kb)

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Tang, Y., Poustovoitov, M., Zhao, K. et al. Structure of a human ASF1a–HIRA complex and insights into specificity of histone chaperone complex assembly. Nat Struct Mol Biol 13, 921–929 (2006). https://doi.org/10.1038/nsmb1147

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