Protein acetylation microarray reveals that NuA4 controls key metabolic target regulating gluconeogenesis.
Histone acetyltransferases (HATs) and histone deacetylases (HDACs) conduct many critical functions through nonhistone substrates in metazoans, but only chromatin-associated nonhistone substrates are known in Saccharomyces cerevisiae. Using yeast proteome microarrays, we identified and validated many nonchromatin substrates of the essential nucleosome acetyltransferase of H4 (NuA4) complex. Among these, acetylation sites (Lys19 and 514) of phosphoenolpyruvate carboxykinase (Pck1p) were determined by tandem mass spectrometry. Acetylation at Lys514 was crucial for enzymatic activity and the ability of yeast cells to grow on nonfermentable carbon sources. Furthermore, Sir2p deacetylated Pck1p both in vitro and in vivo. Loss of Pck1p activity blocked the extension of yeast chronological life span caused by water starvation. In human hepatocellular carcinoma (HepG2) cells, human Pck1 acetylation and glucose production were dependent on TIP60, the human homolog of ESA1. Our findings demonstrate a regulatory function for the NuA4 complex in glucose metabolism and life span by acetylating a critical metabolic enzyme.
Lin
Yu-yi
YY
High Throughput Biology Center, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Lu
Jin-ying
JY
Zhang
Junmei
J
Walter
Wendy
W
Dang
Weiwei
W
Wan
Jun
J
Tao
Sheng-Ce
SC
Qian
Jiang
J
Zhao
Yingming
Y
Boeke
Jef D
JD
Berger
Shelley L
SL
Zhu
Heng
H
D000107:Acetylation; D055785:Gene Knockdown Techniques; D005943:Gluconeogenesis; D005947:Glucose / Q000378:metabolism; D051548:Histone Acetyltransferases / Q000235:genetics / Q000378:metabolism*; D006655:Histone Deacetylases / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D000074443:Lysine Acetyltransferase 5; D046912:Multiprotein Complexes / Q000378:metabolism; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000378:metabolism; D040081:Protein Array Analysis; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D037761:Sirtuins / Q000378:metabolism; D014867:Water / Q000378:metabolism
D047908:Intracellular Signaling Peptides and Proteins; D046912:Multiprotein Complexes; D029701:Saccharomyces cerevisiae Proteins; D014867:Water; D051548:Histone Acetyltransferases; C098489:KAT5 protein, human; D000074443:Lysine Acetyltransferase 5; C504740:NuA4 protein, S cerevisiae; D037761:Sirtuins; D006655:Histone Deacetylases; C521898:PCK1 protein, human; D010729:Phosphoenolpyruvate Carboxykinase (GTP); D005947:Glucose