Structure of the GCN5 histone acetyltransferase bound to a bisubstrate inhibitor

Poux, A.N.; Cebrat, Marek; Kim, Cheol M.; Cole, Philip A.; Marmorstein, Ronen

doi:10.1073/pnas.222373899

Cited by 108 publications

(98 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The structure of HAT domains from several members of the Gcn5/PCAF family has been determined by X-ray crystallography and nuclear magnetic resonance, both alone and in complex with substrates (Clements et al, 1999;Lin et al, 1999;Rojas et al, 1999;Trievel et al, 1999;Yan et al, 2000Yan et al, , 2002Poux et al, 2002;Poux and Marmorstein, 2003). These structures reveal that the A, B and D regions of the GNAT proteins form a central core that is structurally homologous to other GNAT proteins and mediates conserved Ac-CoA interactions ( Figure 1a).…”

Section: Structure Of Nuclear Hatsmentioning

confidence: 99%

“…The observation that the bisubstrate inhibitor Lys-CoA, in which an acetyl bridge is installed between the amine substrate and CoA, is a potent (IC 50 ¼ 500 nM) p300 inhibitor suggested that the enzyme uses a ternary complex mechanism (Sagar et al, 2004). However, neither the H3-CoA-20 nor H4-CoA-20 peptide-CoA conjugates, where CoA is linked analogously to lysines 14 and 8 of the respective histone peptides, are good p300 inhibitors (with IC 50 values above 10 mM), while the H3-CoA-20 is a potent (IC 50 ¼ 360 nM) PCAF inhibitor (Poux et al, 2002). Reciprocally, Lys-CoA is not a good PCAF inhibitor (with an IC 50 of 200 mM).…”

Section: Catalytic Mechanism Of Nuclear Hatsmentioning

confidence: 99%

“…The only direct structural insights into small moleculemediated inhibition of HAT proteins has come from a crystal structure of the Tetrahymena Gcn5 (tGcn5) HAT domain bound to a modified H3-CoA-20 inhibitor (Poux et al, 2002) described above (Figure 2a). The bisubstrate inhibitor used in this study was prepared with an isopropionyl bridge between CoA and peptide to more closely mimic the Ac-CoA-lysine intermediate (Lau et al, 2000b).…”

Section: Structural Basis For Hat Inactivationmentioning

confidence: 99%

See 2 more Smart Citations

Chemistry of acetyl transfer by histone modifying enzymes: structure, mechanism and implications for effector design

2007

View full text Add to dashboard Cite

The post-translational modification of histones plays an important role in chromatin regulation, a process that insures the fidelity of gene expression and other DNA transactions. Of the enzymes that mediate post-translation modification, the histone acetyltransferase (HAT) and histone deacetylase (HDAC) proteins that add and remove acetyl groups to and from target lysine residues within histones, respectively, have been the most extensively studied at both the functional and structural levels. Not surprisingly, the aberrant activity of several of these enzymes have been implicated in human diseases such as cancer and metabolic disorders, thus making them important drug targets. Significant mechanistic insights into the function of HATs and HDACs have come from the X-ray crystal structures of these enzymes both alone and in liganded complexes, along with associated enzymatic and biochemical studies. In this review, we will discuss what we have learned from the structures and related biochemistry of HATs and HDACs and the implications of these findings for the design of protein effectors to regulate gene expression and treat disease.

show abstract

Section: Structure Of Nuclear Hatsmentioning

confidence: 99%

Section: Catalytic Mechanism Of Nuclear Hatsmentioning

confidence: 99%

Section: Structural Basis For Hat Inactivationmentioning

confidence: 99%

See 1 more Smart Citation

Chemistry of acetyl transfer by histone modifying enzymes: structure, mechanism and implications for effector design

2007

View full text Add to dashboard Cite

show abstract

Section: Introductionmentioning

confidence: 99%

“…The kinetic mechanism of Gcn5 has been studied extensively (Tanner et al, 1999;Tanner, Langer, Kim, et al, 2000) and the Gcn5 catalytic domain from several organisms has been crystallized in the presence of various combinations of substrates (Roth et al, 2001;Poux et al, 2002). The active site of Gcn5 contains two grooves where acetyl-CoA and peptide bind, which intersect near the β-mercaptoethylamine moiety of coenzyme A and the target lysine Poux et al, 2002). The ternary complex between Gcn5, acetyl-CoA, and peptide forms through a fully ordered mechanism (Tanner, Langer, Kim, et al, 2000), as binding to acetyl-CoA brings about a structural rearrangement that widens the peptide-binding groove within the Gcn5 active site (Clements et al, 1999;Trievel et al, 1999;Rojas et al, 1999;Lin et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

Structural basis for acyl group discrimination by human Gcn5L2

Ringel

Wolberger

2016

Preprint

View full text Add to dashboard Cite

Gcn5 is a conserved acetyltransferase that regulates transcription by acetylating the Nterminal tails of histones. Motivated by recent studies identifying a chemically diverse array of lysine acyl modifications in vivo, we examined the acyl chain specificity of the acetyltransferase, human Gcn5 (Gcn5L2). Whereas Gcn5L2 robustly catalyzes lysine acetylation, the acyltransferase activity of Gcn5L2 gets progressively weaker with increasing acyl chain length. To understand how Gcn5 discriminates between different acyl-CoA molecules, we determined structures of the catalytic domain of human Gcn5L2 bound to propionyl-CoA and butyryl-CoA. Although the active site of Gcn5L2 can accommodate propionyl-CoA and butyryl-CoA without major structural rearrangements, butyryl-CoA adopts a conformation incompatible with catalysis that obstructs the path of the incoming lysine residue and acts as a competitive inhibitor for Gcn5L2 versus acetyl-CoA. These structures demonstrate how Gcn5L2 discriminates between acyl chain donors and explain why Gcn5L2 has weak activity for acyl moieties that are larger than an acetyl group.

show abstract

Generating enzyme and radical‐mediated bisubstrates as tools for investigating Gcn5‐related N‐acetyltransferases

et al. 2017

View full text Add to dashboard Cite

Gcn5-related N-acetyltransferases (GNATs) are found in all kingdoms of life and catalyze important acyl transfer reactions in diverse cellular processes. While many 3D structures of GNATs have been determined, most do not contain acceptor substrates in their active sites. To expand upon existing crystallographic strategies for improving acceptor-bound GNAT structures, we synthesized peptide substrate analogs and reacted them with CoA in PA4794 protein crystals. We found two separate mechanisms for bisubstrate formation: 1) a novel X-ray induced radical-mediated alkylation of CoA with an alkene peptide, and 2) direct alkylation of CoA with a halogenated peptide. Our approach is widely applicable across the GNAT superfamily and can be used to improve the success rate of obtaining liganded structures of other acyltransferases.

show abstract

Structure of the GCN5 histone acetyltransferase bound to a bisubstrate inhibitor

Cited by 108 publications

References 27 publications

Chemistry of acetyl transfer by histone modifying enzymes: structure, mechanism and implications for effector design

Chemistry of acetyl transfer by histone modifying enzymes: structure, mechanism and implications for effector design

Structural basis for acyl group discrimination by human Gcn5L2

Generating enzyme and radical‐mediated bisubstrates as tools for investigating Gcn5‐related N‐acetyltransferases

Contact Info

Product

Resources

About