Methylation of Histone H3 Mediates the Association of the NuA3 Histone Acetyltransferase with Chromatin

Abstract

The SAS3-dependent NuA3 histone acetyltransferase complex was originally identified on the basis of its ability to acetylate histone H3 in vitro. Whether NuA3 is capable of acetylating histones in vivo, or how the complex is targeted to the nucleosomes that it modifies, was unknown. To address this question, we asked whether NuA3 is associated with chromatin in vivo and how this association is regulated. With a chromatin pulldown assay, we found that NuA3 interacts with the histone H3 amino-terminal tail, and loss of the H3 tail recapitulates phenotypes associated with loss of SAS3. Moreover, mutation of histone H3 lysine 14, the preferred site of acetylation by NuA3 in vitro, phenocopies a unique sas3Δ phenotype, suggesting that modification of this residue is important for NuA3 function. The interaction of NuA3 with chromatin is dependent on the Set1p and Set2p histone methyltransferases, as well as their substrates, histone H3 lysines 4 and 36, respectively. These results confirm that NuA3 is functioning as a histone acetyltransferase in vivo and that histone H3 methylation provides a mark for the recruitment of NuA3 to nucleosomes.

Support for this work was provided by a grant to L.H. from the Cancer Research Institute of the Canadian Institute for Health Research. L.H. is a scholar of the Michael Smith Foundation for Health Research.

We gratefully acknowledge the valuable comments provided by Jacques Côté, Jerry Workman, and members of the Molecular Epigenetics Group of the Life Sciences Institute at the University of British Columbia. We are also grateful to Fred Winston and Jasper Rine for providing yeast strains and plasmids.