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Fungal Rtt109 histone acetyltransferase is an unexpected structural homolog of metazoan p300/CBP

Nature Structural & Molecular Biology volume 15, pages 738–745 (2008)Cite this article

A Corrigendum to this article was published on 01 September 2008

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Abstract

Rtt109, also known as KAT11, is a recently characterized fungal-specific histone acetyltransferase (HAT) that modifies histone H3 lysine 56 (H3K56) to promote genome stability. Rtt109 does not show sequence conservation with other known HATs and depends on association with either of two histone chaperones, Asf1 or Vps75, for HAT activity. Here we report the X-ray crystal structure of an Rtt109–acetyl coenzyme A complex and carry out structure-based mutagenesis, combined with in vitro biochemical studies of the Rtt109–Vps75 complex and studies of Rtt109 function in vivo. The Rtt109 structure reveals noteworthy homology to the metazoan p300/CBP HAT domain but exhibits functional divergence, including atypical catalytic properties and mode of cofactor regulation. The structure reveals a buried autoacetylated lysine residue that we show is also acetylated in the Rtt109 protein purified from yeast cells. Implications for understanding histone substrate and chaperone binding by Rtt109 are discussed.

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Figure 1: Overall structure of the Rtt109–acetyl-CoA complex and its relationship to p300.
Figure 2: Detailed view of the Rtt109–acetyl-CoA structure.
Figure 3: Structure-guided mutagenesis of the active site.
Figure 4: H3K56 acetylation and sensitivity to genotoxic agents in S. cerevisiae strains expressing wild-type and mutant forms of Rtt109.
Figure 5: Rtt109 autoacetylation at Lys290.

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  • 30 July 2008

    In the version of this article initially published, the author name Verreault was mistakenly spelled as Verrault. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank M. Allaire for assistance with crystallographic data collection and K. Speicher, T. Beer and D. Speicher in the Wistar Institute Proteomics Core Facility for performing the in-gel digestion, LC-MS/MS, database searching and analysis on the recombinant Rtt109 proteins. This work was supported by US National Institutes of Health grants to R.M. and P.A.C. 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. Research in the A.V. and P.T. laboratories is funded by the Canadian Institutes for Health Research (CIHR) operating funds. The Institute for Research in Immunology and Cancer receives infrastructure funds from Fonds de Recherche en Sante du Quebec and a multi-resources facility grant from CIHR.

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Authors and Affiliations

  1. Program in Gene Expression and Regulation, The Wistar Institute, 3601 Spruce Street, Philadelphia, 19104, Pennsylvania, USA

    Yong Tang, Katrina Meeth, Eva Jiang & Ronen Marmorstein

  2. Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, 21205, Maryland, USA

    Marc A Holbert & Philip A Cole

  3. Institute for Research in Immunology and Cancer, Université de Montréal, PO Box 6128, Succursale Centre-Ville, Montréal, H3C 3J7, QC, Canada

    Hugo Wurtele, Walter Rocha, Marlene Gharib, Pierre Thibault & Alain Verreault

  4. Department of Chemistry, University of Pennsylvania, 231 South 34th Street, Philadelphia, 19104, Pennsylvania, USA

    Ronen Marmorstein

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Contributions

Y.T., M.A.H., H.W., K.M., W.R. and M.G. designed and carried out experiments reported in the manuscript, and prepared manuscript figures and text; E.J. carried out preliminary studies that led to experiments reported in the manuscript; P.T., A.V., P.A.C. 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 Ronen Marmorstein.

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Tang, Y., Holbert, M., Wurtele, H. et al. Fungal Rtt109 histone acetyltransferase is an unexpected structural homolog of metazoan p300/CBP. Nat Struct Mol Biol 15, 738–745 (2008). https://doi.org/10.1038/nsmb.1448

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