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Lactate, a product of glycolytic metabolism, inhibits histone deacetylase activity and promotes changes in gene expression.

Latham T, Mackay L, Sproul D, Karim M, Culley J, Harrison DJ, Hayward L, Langridge-Smith P, Gilbert N, Ramsahoye BH - Nucleic Acids Res. (2012)

Bottom Line: Lactate is a relatively weak inhibitor (IC(50) 40 mM) compared to the established inhibitors trichostatin A and butyrate, but the genes deregulated overlap significantly with those affected by low concentrations of the more potent inhibitors.HDAC inhibition causes significant up and downregulation of genes, but genes that are associated with HDAC proteins are more likely to be upregulated and less likely to be downregulated than would be expected.Our results suggest that the primary effect of HDAC inhibition by endogenous short-chain fatty acids like lactate is to promote gene expression at genes associated with HDAC proteins.

View Article: PubMed Central - PubMed

Affiliation: Institute of Genetics and Molecular Medicine, University of Edinburgh, Crewe Road South, Edinburgh, EH4 2XR, SIRCAMS, UK.

ABSTRACT
Chemical inhibitors of histone deacetylase (HDAC) activity are used as experimental tools to induce histone hyperacetylation and deregulate gene transcription, but it is not known whether the inhibition of HDACs plays any part in the normal physiological regulation of transcription. Using both in vitro and in vivo assays, we show that lactate, which accumulates when glycolysis exceeds the cell's aerobic metabolic capacity, is an endogenous HDAC inhibitor, deregulating transcription in an HDAC-dependent manner. Lactate is a relatively weak inhibitor (IC(50) 40 mM) compared to the established inhibitors trichostatin A and butyrate, but the genes deregulated overlap significantly with those affected by low concentrations of the more potent inhibitors. HDAC inhibition causes significant up and downregulation of genes, but genes that are associated with HDAC proteins are more likely to be upregulated and less likely to be downregulated than would be expected. Our results suggest that the primary effect of HDAC inhibition by endogenous short-chain fatty acids like lactate is to promote gene expression at genes associated with HDAC proteins. Therefore, we propose that lactate may be an important transcriptional regulator, linking the metabolic state of the cell to gene transcription.

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Exhausted tissue culture medium exhibits HDAC inhibitory activity due to the accumulation of l-lactate and increasing acidity. (A) In vitro HDAC assays to test the HDAC-inhibitory activities of fresh (open circles) and used tissue culture medium (black diamonds). All activities are percentages of the reaction buffer control. The media contributed 40% by volume to the total HDAC reaction mix. The pH values of the reaction mixtures are the same throughout (range pH 8.09–8.12). (B) NMR spectroscopy of complete tissue culture medium at various times in culture following a medium change. The control profile is of fresh medium spiked with an l-lactate standard. (C) Lactate measurements in fresh and used tissue culture medium. (D) l-Lactate, d-lactate and low-pH inhibit histone deacetylase activity in vitro. The graph shows HDAC activity in the presence of increasing concentrations of NaCl (control, filled circles), butyrate (open circles), TSA (black crosses), l-lactate (black diamonds), d-lactate (black triangles) and pyruvate (black squares). (E) In vitro HDAC activity is inhibited at low pH.
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gks066-F2: Exhausted tissue culture medium exhibits HDAC inhibitory activity due to the accumulation of l-lactate and increasing acidity. (A) In vitro HDAC assays to test the HDAC-inhibitory activities of fresh (open circles) and used tissue culture medium (black diamonds). All activities are percentages of the reaction buffer control. The media contributed 40% by volume to the total HDAC reaction mix. The pH values of the reaction mixtures are the same throughout (range pH 8.09–8.12). (B) NMR spectroscopy of complete tissue culture medium at various times in culture following a medium change. The control profile is of fresh medium spiked with an l-lactate standard. (C) Lactate measurements in fresh and used tissue culture medium. (D) l-Lactate, d-lactate and low-pH inhibit histone deacetylase activity in vitro. The graph shows HDAC activity in the presence of increasing concentrations of NaCl (control, filled circles), butyrate (open circles), TSA (black crosses), l-lactate (black diamonds), d-lactate (black triangles) and pyruvate (black squares). (E) In vitro HDAC activity is inhibited at low pH.

Mentions: We looked for inhibitory activity against type I and type II HDACs in Hela cell nuclei using a fluorescent HDAC substrate (Biomol HDAC assay, see ‘Materials and Methods’ section). We found that HDAC inhibition was proportionate to the amount of exhausted medium in the assay (Figure 2A). We then looked for compounds that might be responsible for HDAC inhibition by screening used medium by NMR spectroscopy. We identified a compound giving a double peak at a resonance of 1.3 ppm (Figure 2B). This signature is characteristic of lactate. We confirmed the compound’s identity by replicating the signature using fresh medium spiked with an l-lactate standard (Figure 2B, control).Figure 2.


Lactate, a product of glycolytic metabolism, inhibits histone deacetylase activity and promotes changes in gene expression.

Latham T, Mackay L, Sproul D, Karim M, Culley J, Harrison DJ, Hayward L, Langridge-Smith P, Gilbert N, Ramsahoye BH - Nucleic Acids Res. (2012)

Exhausted tissue culture medium exhibits HDAC inhibitory activity due to the accumulation of l-lactate and increasing acidity. (A) In vitro HDAC assays to test the HDAC-inhibitory activities of fresh (open circles) and used tissue culture medium (black diamonds). All activities are percentages of the reaction buffer control. The media contributed 40% by volume to the total HDAC reaction mix. The pH values of the reaction mixtures are the same throughout (range pH 8.09–8.12). (B) NMR spectroscopy of complete tissue culture medium at various times in culture following a medium change. The control profile is of fresh medium spiked with an l-lactate standard. (C) Lactate measurements in fresh and used tissue culture medium. (D) l-Lactate, d-lactate and low-pH inhibit histone deacetylase activity in vitro. The graph shows HDAC activity in the presence of increasing concentrations of NaCl (control, filled circles), butyrate (open circles), TSA (black crosses), l-lactate (black diamonds), d-lactate (black triangles) and pyruvate (black squares). (E) In vitro HDAC activity is inhibited at low pH.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3367171&req=5

gks066-F2: Exhausted tissue culture medium exhibits HDAC inhibitory activity due to the accumulation of l-lactate and increasing acidity. (A) In vitro HDAC assays to test the HDAC-inhibitory activities of fresh (open circles) and used tissue culture medium (black diamonds). All activities are percentages of the reaction buffer control. The media contributed 40% by volume to the total HDAC reaction mix. The pH values of the reaction mixtures are the same throughout (range pH 8.09–8.12). (B) NMR spectroscopy of complete tissue culture medium at various times in culture following a medium change. The control profile is of fresh medium spiked with an l-lactate standard. (C) Lactate measurements in fresh and used tissue culture medium. (D) l-Lactate, d-lactate and low-pH inhibit histone deacetylase activity in vitro. The graph shows HDAC activity in the presence of increasing concentrations of NaCl (control, filled circles), butyrate (open circles), TSA (black crosses), l-lactate (black diamonds), d-lactate (black triangles) and pyruvate (black squares). (E) In vitro HDAC activity is inhibited at low pH.
Mentions: We looked for inhibitory activity against type I and type II HDACs in Hela cell nuclei using a fluorescent HDAC substrate (Biomol HDAC assay, see ‘Materials and Methods’ section). We found that HDAC inhibition was proportionate to the amount of exhausted medium in the assay (Figure 2A). We then looked for compounds that might be responsible for HDAC inhibition by screening used medium by NMR spectroscopy. We identified a compound giving a double peak at a resonance of 1.3 ppm (Figure 2B). This signature is characteristic of lactate. We confirmed the compound’s identity by replicating the signature using fresh medium spiked with an l-lactate standard (Figure 2B, control).Figure 2.

Bottom Line: Lactate is a relatively weak inhibitor (IC(50) 40 mM) compared to the established inhibitors trichostatin A and butyrate, but the genes deregulated overlap significantly with those affected by low concentrations of the more potent inhibitors.HDAC inhibition causes significant up and downregulation of genes, but genes that are associated with HDAC proteins are more likely to be upregulated and less likely to be downregulated than would be expected.Our results suggest that the primary effect of HDAC inhibition by endogenous short-chain fatty acids like lactate is to promote gene expression at genes associated with HDAC proteins.

View Article: PubMed Central - PubMed

Affiliation: Institute of Genetics and Molecular Medicine, University of Edinburgh, Crewe Road South, Edinburgh, EH4 2XR, SIRCAMS, UK.

ABSTRACT
Chemical inhibitors of histone deacetylase (HDAC) activity are used as experimental tools to induce histone hyperacetylation and deregulate gene transcription, but it is not known whether the inhibition of HDACs plays any part in the normal physiological regulation of transcription. Using both in vitro and in vivo assays, we show that lactate, which accumulates when glycolysis exceeds the cell's aerobic metabolic capacity, is an endogenous HDAC inhibitor, deregulating transcription in an HDAC-dependent manner. Lactate is a relatively weak inhibitor (IC(50) 40 mM) compared to the established inhibitors trichostatin A and butyrate, but the genes deregulated overlap significantly with those affected by low concentrations of the more potent inhibitors. HDAC inhibition causes significant up and downregulation of genes, but genes that are associated with HDAC proteins are more likely to be upregulated and less likely to be downregulated than would be expected. Our results suggest that the primary effect of HDAC inhibition by endogenous short-chain fatty acids like lactate is to promote gene expression at genes associated with HDAC proteins. Therefore, we propose that lactate may be an important transcriptional regulator, linking the metabolic state of the cell to gene transcription.

Show MeSH
Related in: MedlinePlus