MYST protein acetyltransferase activity requires active site lysine autoacetylation.
The MYST protein lysine acetyltransferases are evolutionarily conserved throughout eukaryotes and acetylate proteins to regulate diverse biological processes including gene regulation, DNA repair, cell-cycle regulation, stem cell homeostasis and development. Here, we demonstrate that MYST protein acetyltransferase activity requires active site lysine autoacetylation. The X-ray crystal structures of yeast Esa1 (yEsa1/KAT5) bound to a bisubstrate H4K16CoA inhibitor and human MOF (hMOF/KAT8/MYST1) reveal that they are autoacetylated at a strictly conserved lysine residue in MYST proteins (yEsa1-K262 and hMOF-K274) in the enzyme active site. The structure of hMOF also shows partial occupancy of K274 in the unacetylated form, revealing that the side chain reorients to a position that engages the catalytic glutamate residue and would block cognate protein substrate binding. Consistent with the structural findings, we present mass spectrometry data and biochemical experiments to demonstrate that this lysine autoacetylation on yEsa1, hMOF and its yeast orthologue, ySas2 (KAT8) occurs in solution and is required for acetylation and protein substrate binding in vitro. We also show that this autoacetylation occurs in vivo and is required for the cellular functions of these MYST proteins. These findings provide an avenue for the autoposttranslational regulation of MYST proteins that is distinct from other acetyltransferases but draws similarities to the phosphoregulation of protein kinases.
Gene Expression and Regulation Program, The Wistar Institute, University of Pennsylvania, Philadelphia, PA 19104, USA.
Hua
H
Yuan
Dorine
D
Rossetto
Hestia
H
Mellert
Weiwei
W
Dang
Madhusudan
M
Srinivasan
Jamel
J
Johnson
Santosh
S
Hodawadekar
Emily C
EC
Ding
Kaye
K
Speicher
Nebiyu
N
Abshiru
Rocco
R
Perry
Jiang
J
Wu
Chao
C
Yang
Y George
YG
Zheng
David W
DW
Speicher
Pierre
P
Thibault
Alain
A
Verreault
F Bradley
FB
Johnson
Shelley L
SL
Berger
Rolf
R
Sternglanz
Steven B
SB
McMahon
Jacques
J
Côté
Ronen
R
Marmorstein
D000107:Acetylation; D001665:Binding Sites; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D051548:Histone Acetyltransferases / Q000235:genetics / Q000378:metabolism*; D006657:Histones / Q000378:metabolism; D006801:Humans; D008239:Lysine / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism
D006657:Histones; D029701:Saccharomyces cerevisiae Proteins; C496551:Esa1 protein, S cerevisiae; D051548:Histone Acetyltransferases; C496552:KAT8 protein, human; C469266:Sas2 protein, S cerevisiae; D008239:Lysine