Impact of ATP-citrate lyase catalytic activity and serine 455 phosphorylation on histone acetylation and inflammatory responses in human monocytic THP-1 cells
ATP-citrate lyase (ACLY) is a key enzyme provoking metabolic and epigenetic gene regulation. Molecularly, these functions are exerted by the provision of acetyl-coenzyme A, which is then used as a substrate for de novo lipogenesis or as an acetyl-group donor in acetylation reactions. It has been dem...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-11-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.906127/full |
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| author | Monica Dominguez Verena Truemper Ana Carolina Mota Bernhard Brüne Bernhard Brüne Bernhard Brüne Bernhard Brüne Dmitry Namgaladze |
| author_facet | Monica Dominguez Verena Truemper Ana Carolina Mota Bernhard Brüne Bernhard Brüne Bernhard Brüne Bernhard Brüne Dmitry Namgaladze |
| author_sort | Monica Dominguez |
| collection | DOAJ |
| description | ATP-citrate lyase (ACLY) is a key enzyme provoking metabolic and epigenetic gene regulation. Molecularly, these functions are exerted by the provision of acetyl-coenzyme A, which is then used as a substrate for de novo lipogenesis or as an acetyl-group donor in acetylation reactions. It has been demonstrated that ACLY activity can be positively regulated via phosphorylation at serine 455 by Akt and protein kinase A. Nonetheless, the impact of phosphorylation on ACLY function in human myeloid cells is poorly understood. In this study we reconstituted ACLY knockout human monocytic THP-1 cells with a wild type ACLY as well as catalytically inactive H760A, and phosphorylation-deficient S455A mutants. Using these cell lines, we determined the impact of ACLY activity and phosphorylation on histone acetylation and pro-inflammatory gene expression in response to lipopolysaccharide (LPS). Our results show that ACLY serine 455 phosphorylation does not influence the proper enzymatic function of ACLY, since both, wild type ACLY and phosphorylation-deficient mutant, exhibited increased cell growth and histone acetylation as compared to cells with a loss of ACLY activity. Transcriptome analysis revealed enhanced expression of pro-inflammatory and interferon response genes in ACLY knockout and H760A THP-1 cells under unstimulated or LPS-treated conditions. At the same time, S455A ACLY-expressing cells showed a phenotype very similar to wild type cells. Contrary to ACLY knockout, pharmacological inhibition of ACLY in THP-1 cells or in primary human macrophages does not enhance LPS-triggered pro-inflammatory gene expression. Our data thus suggest that ACLY retains functionality in the absence of Akt/PKA-mediated phosphorylation in human myeloid cells. Furthermore, loss of ACLY activity may elicit long-term adaptive mechanisms, increasing inflammatory responses. |
| first_indexed | 2024-04-11T17:33:44Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-11T17:33:44Z |
| publishDate | 2022-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-a6acc1ea8bf649efad6d17caefe54cea2022-12-22T04:11:56ZengFrontiers Media S.A.Frontiers in Immunology1664-32242022-11-011310.3389/fimmu.2022.906127906127Impact of ATP-citrate lyase catalytic activity and serine 455 phosphorylation on histone acetylation and inflammatory responses in human monocytic THP-1 cellsMonica Dominguez0Verena Truemper1Ana Carolina Mota2Bernhard Brüne3Bernhard Brüne4Bernhard Brüne5Bernhard Brüne6Dmitry Namgaladze7Institute of Biochemistry I, Faculty of Medicine, Goethe-University Frankfurt, Frankfurt, GermanyInstitute of Biochemistry I, Faculty of Medicine, Goethe-University Frankfurt, Frankfurt, GermanyInstitute of Biochemistry I, Faculty of Medicine, Goethe-University Frankfurt, Frankfurt, GermanyInstitute of Biochemistry I, Faculty of Medicine, Goethe-University Frankfurt, Frankfurt, GermanyFraunhofer Institute for Translational Medicine and Pharmacology (ITMP), Frankfurt, GermanyGerman Cancer Consortium (DKTK), Partner Site Frankfurt, Frankfurt, GermanyFrankfurt Cancer Institute, Goethe-University Frankfurt, Frankfurt, GermanyInstitute of Biochemistry I, Faculty of Medicine, Goethe-University Frankfurt, Frankfurt, GermanyATP-citrate lyase (ACLY) is a key enzyme provoking metabolic and epigenetic gene regulation. Molecularly, these functions are exerted by the provision of acetyl-coenzyme A, which is then used as a substrate for de novo lipogenesis or as an acetyl-group donor in acetylation reactions. It has been demonstrated that ACLY activity can be positively regulated via phosphorylation at serine 455 by Akt and protein kinase A. Nonetheless, the impact of phosphorylation on ACLY function in human myeloid cells is poorly understood. In this study we reconstituted ACLY knockout human monocytic THP-1 cells with a wild type ACLY as well as catalytically inactive H760A, and phosphorylation-deficient S455A mutants. Using these cell lines, we determined the impact of ACLY activity and phosphorylation on histone acetylation and pro-inflammatory gene expression in response to lipopolysaccharide (LPS). Our results show that ACLY serine 455 phosphorylation does not influence the proper enzymatic function of ACLY, since both, wild type ACLY and phosphorylation-deficient mutant, exhibited increased cell growth and histone acetylation as compared to cells with a loss of ACLY activity. Transcriptome analysis revealed enhanced expression of pro-inflammatory and interferon response genes in ACLY knockout and H760A THP-1 cells under unstimulated or LPS-treated conditions. At the same time, S455A ACLY-expressing cells showed a phenotype very similar to wild type cells. Contrary to ACLY knockout, pharmacological inhibition of ACLY in THP-1 cells or in primary human macrophages does not enhance LPS-triggered pro-inflammatory gene expression. Our data thus suggest that ACLY retains functionality in the absence of Akt/PKA-mediated phosphorylation in human myeloid cells. Furthermore, loss of ACLY activity may elicit long-term adaptive mechanisms, increasing inflammatory responses.https://www.frontiersin.org/articles/10.3389/fimmu.2022.906127/fullATP-citrate lyasehistone acetylationmacrophagesmetabolisminflammation |
| spellingShingle | Monica Dominguez Verena Truemper Ana Carolina Mota Bernhard Brüne Bernhard Brüne Bernhard Brüne Bernhard Brüne Dmitry Namgaladze Impact of ATP-citrate lyase catalytic activity and serine 455 phosphorylation on histone acetylation and inflammatory responses in human monocytic THP-1 cells Frontiers in Immunology ATP-citrate lyase histone acetylation macrophages metabolism inflammation |
| title | Impact of ATP-citrate lyase catalytic activity and serine 455 phosphorylation on histone acetylation and inflammatory responses in human monocytic THP-1 cells |
| title_full | Impact of ATP-citrate lyase catalytic activity and serine 455 phosphorylation on histone acetylation and inflammatory responses in human monocytic THP-1 cells |
| title_fullStr | Impact of ATP-citrate lyase catalytic activity and serine 455 phosphorylation on histone acetylation and inflammatory responses in human monocytic THP-1 cells |
| title_full_unstemmed | Impact of ATP-citrate lyase catalytic activity and serine 455 phosphorylation on histone acetylation and inflammatory responses in human monocytic THP-1 cells |
| title_short | Impact of ATP-citrate lyase catalytic activity and serine 455 phosphorylation on histone acetylation and inflammatory responses in human monocytic THP-1 cells |
| title_sort | impact of atp citrate lyase catalytic activity and serine 455 phosphorylation on histone acetylation and inflammatory responses in human monocytic thp 1 cells |
| topic | ATP-citrate lyase histone acetylation macrophages metabolism inflammation |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.906127/full |
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