Catalytic activation of histone acetyltransferase Rtt109 by a histone chaperone

Department of Biomolecular Chemistry, School of Medicine and Public Health, University of Wisconsin, 1300 University Avenue, Madison, WI 53706, USA.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 11/2010; 107(47):20275-80. DOI: 10.1073/pnas.1009860107
Source: PubMed


Most histone acetyltransferases (HATs) function as multisubunit complexes in which accessory proteins regulate substrate specificity and catalytic efficiency. Rtt109 is a particularly interesting example of a HAT whose specificity and catalytic activity require association with either of two histone chaperones, Vps75 or Asf1. Here, we utilize biochemical, structural, and genetic analyses to provide the detailed molecular mechanism for activation of a HAT (Rtt109) by its activating subunit Vps75. The rate-determining step of the activated complex is the transfer of the acetyl group from acetyl CoA to the acceptor lysine residue. Vps75 stimulates catalysis (> 250-fold), not by contributing a catalytic base, but by stabilizing the catalytically active conformation of Rtt109. To provide structural insight into the functional complex, we produced a molecular model of Rtt109-Vps75 based on X-ray diffraction of crystals of the complex. This model reveals distinct negative electrostatic surfaces on an Rtt109 molecule that interface with complementary electropositive ends of a symmetrical Vps75 dimer. Rtt109 variants with interface point substitutions lack the ability to be fully activated by Vps75, and one such variant displayed impaired Vps75-dependent histone acetylation functions in yeast, yet these variants showed no adverse effect on Asf1-dependent Rtt109 activities in vitro and in vivo. Finally, we provide evidence for a molecular model in which a 12 complex of Rtt109-Vps75 acetylates a heterodimer of H3-H4. The activation mechanism of Rtt109-Vps75 provides a valuable framework for understanding the molecular regulation of HATs within multisubunit complexes.

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    • "In Saccharomyces cerevisiae, the transfer of the acetyl group to H3K56 is catalysed by HAT Rtt109 in complex with the H3-H4 dimer (Fillingham et al., 2008; Kolonko et al., 2010). Because of the lack of Rtt109 "
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    • "In a cellular context, a proportion of Vps75 is found stably associated with the histone acetyltransferase Rtt109. Interestingly, Rtt109 has been observed to form ring-shaped complexes with Vps75 (10,18,19). However, there appears to be some plasticity in the interface between Vps75 and Rtt109 as both 2:1 and 2:2 Vps75:Rtt109 complexes have been reported. "
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    • "By contrast, early characterization of Vps75 demonstrated that this chaperone can bind tetrameric (H3-H4) 2 in vitro (Selth and Svejstrup, 2007). This observation was recently confirmed by structural and biophysical analyses (Bowman et al., 2011; Tang et al., 2011), but it has also been disputed (Kolonko et al., 2010). While this disagreement remains to be rectified, it appears that at least some chaperones are capable of transporting tetrameric histones. "
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