Dataset Information

Human KAT2A/B-acetylome MudPIT

ABSTRACT: Comprehensive identification of acetylated proteins targeted by the lysine acetyltransferases KAT2A (GCN5) and KAT2B (PCAF), in human cells. KAT2A/B-dependent acetylated proteins were identified by comparing acetylated proteins in whole cell extract of control cells and cells in which KAT2A and KAT2B were simultaneously knocked down. The identification of acetylated proteins was performed using an enrichment-free approach. In order to do so, while facing the challenge of identifying low abundant acetylated peptides within a complex sample mixture, we have screened acetylated peptides starting from a large amount of material, equivalent to 50ug of protein digest (maximum loading capacity of our analytical chromatographic column). By analysing such a large amount of peptides, the level of acetylated peptides was then increased and thus acetylated peptides became easily detectable by mass spectrometry, without the need for enrichment.

INSTRUMENT(S): LTQ

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Mathilde Joint

LAB HEAD: Laszlo TORA

PROVIDER: PXD004669 | Pride | 2016-12-12

REPOSITORIES: Pride

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Publications

KAT2A/KAT2B-targeted acetylome reveals a role for PLK4 acetylation in preventing centrosome amplification.

Fournier Marjorie M Orpinell Meritxell M Grauffel Cédric C Scheer Elisabeth E Garnier Jean-Marie JM Ye Tao T Chavant Virginie V Joint Mathilde M Esashi Fumiko F Dejaegere Annick A Gönczy Pierre P Tora László L

Nature communications 20161031

Lysine acetylation is a widespread post-translational modification regulating various biological processes. To characterize cellular functions of the human lysine acetyltransferases KAT2A (GCN5) and KAT2B (PCAF), we determined their acetylome by shotgun proteomics. One of the newly identified KAT2A/2B substrate is polo-like kinase 4 (PLK4), a key regulator of centrosome duplication. We demonstrate that KAT2A/2B acetylate the PLK4 kinase domain on residues K45 and K46. Molecular dynamics modellin ...[more]

PMID: 27796307

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Project description:During fasting, increases in circulating pancreatic glucagon maintain glucose balance by up-regulating hepatic gluconeogenesis. Triggering of the cAMP pathway stimulates the gluconeogenic program through the phosphorylation of CREB and via the de-phosphorylation of the CREB coactivator CRTC2. Hormonal and nutrient signals are also thought to modulate gluconeogenic genes by promoting epigenetic changes that facilitate assembly of the transcriptional machinery, although the nature of these modifications is unclear. Here we show that histone H3 acetylation at Lys 9 (H3K9Ac) is elevated over gluconeogenic genes during fasting and in diabetes, where it contributes to increases in hepatic glucose production. Following its dephosphorylation, CRTC2 promoted increases in H3K9Ac by mediating the recruitment of the lysine acetyltransferase 2B (KAT2B) and WD repeat-containing protein 5 (WDR5), a core subunit of histone methyltransferase (HMT) complexes. In turn, KAT2B and WDR5 stimulated the gluconeogenic program through a self-reinforcing cycle whereby increases in H3K9Ac further potentiated CRTC2 occupancy at CREB binding sites. Breaking this cycle, by depletion of KAT2B or WDR5, decreased gluconeogenic gene expression. As administration of a small molecule KAT2B antagonist lowered circulating blood glucose concentrations in insulin resistance, our results demonstrate how this enzyme may be a useful target for diabetes treatment. A subset of cAMP responsive genes depend on specific recruitment of KAT2B (pcaf), which in concert with WDR5 acetylates H3K9. By selectively depleting hepatocytes for KAT2B or WDR5 prior to glucagon stimulation we explore, which genes rely on KAT2B and WDR5 activity. mKAT2B or mWDR5 were knocked down in primary mouse hepatocytes using adenoviral transduction with appropriate shRNAs. Control cells were transduced with a non-specific (NS) shRNA. 72 hours post transduction some cells were stimulated for 90 minutes with 100nM glucagon and others with PBS. Total RNA was purified and subjected to micro-RNA analysis. All samples are pools of RNA from three sepearate dishes. One replicate is included for most samples.

Dataset Information

Human KAT2A/B-acetylome MudPIT

Publications

KAT2A/KAT2B-targeted acetylome reveals a role for PLK4 acetylation in preventing centrosome amplification.

Similar Datasets

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