Abstract
Rtt109 is a histone acetyltransferase that requires a histone chaperone for the acetylation of histone 3 at lysine 56 (H3K56). Rtt109 forms a complex with the chaperone Vps75 in vivo and is implicated in DNA replication and repair. Here we show that both Rtt109 and Vps75 bind histones with high affinity, but only the complex is efficient for catalysis. The C-terminal acidic domain of Vps75 contributes to activation of Rtt109 and is necessary for in vivo functionality of Vps75, but it is not required for interaction with either Rtt109 or histones. We demonstrate that Vps75 is a structural homolog of yeast Nap1 by solving its crystal structure. Nap1 and Vps75 interact with histones and Rtt109 with comparable affinities. However, only Vps75 stimulates Rtt109 enzymatic activity. Our data highlight the functional specificity of Vps75 in Rtt109 activation.
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Acknowledgements
We thank S. McBryant for help with analytical ultracentrifugation, N.J. Krogan, D.A. Goldstrohm and C.A. Radebaugh for discussion, and K. Brown for critical reading of the manuscript. We also thank J. Nix at MBC 4.2.2., Advanced Light Source, for data collection. This work was funded by the US National Institutes of Health (NIH) grant GM067777 to K.L. and by NIH grant GM056884 to L.A.S. Y.-J.P., A.J.A. and K.L. are supported by the Howard Hughes Medical Institute.
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Y.-.J.P. performed crystallography, gel shifts, enzymatic assays and initial yeast genetics studies; K.B.S. performed in vivo experiments; A.J.A. performed quantitative protein-protein interaction assays, MS and enzymatic assays; L.A.S. performed planning and advice for in vivo experiments; K.L. and Y.-J.P. planned and supervised the structural and biochemical experimental sections.
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Park, YJ., Sudhoff, K., Andrews, A. et al. Histone chaperone specificity in Rtt109 activation. Nat Struct Mol Biol 15, 957–964 (2008). https://doi.org/10.1038/nsmb.1480
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DOI: https://doi.org/10.1038/nsmb.1480
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