HIV latency is reversed by ACSS2-driven histone crotonylation
Guochun Jiang, … , Joseph K. Wong, Satya Dandekar
Guochun Jiang, … , Joseph K. Wong, Satya Dandekar
Published February 19, 2018
Citation Information: J Clin Invest. 2018;128(3):1190-1198. https://doi.org/10.1172/JCI98071.
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Research Article AIDS/HIV Infectious disease

HIV latency is reversed by ACSS2-driven histone crotonylation

Abstract

Eradication of HIV-1 (HIV) is hindered by stable viral reservoirs. Viral latency is epigenetically regulated. While the effects of histone acetylation and methylation at the HIV long-terminal repeat (LTR) have been described, our knowledge of the proviral epigenetic landscape is incomplete. We report that a previously unrecognized epigenetic modification of the HIV LTR, histone crotonylation, is a regulator of HIV latency. Reactivation of latent HIV was achieved following the induction of histone crotonylation through increased expression of the crotonyl-CoA–producing enzyme acyl-CoA synthetase short-chain family member 2 (ACSS2). This reprogrammed the local chromatin at the HIV LTR through increased histone acetylation and reduced histone methylation. Pharmacologic inhibition or siRNA knockdown of ACSS2 diminished histone crotonylation–induced HIV replication and reactivation. ACSS2 induction was highly synergistic in combination with either a protein kinase C agonist (PEP005) or a histone deacetylase inhibitor (vorinostat) in reactivating latent HIV. In the SIV-infected nonhuman primate model of AIDS, the expression of ACSS2 was significantly induced in intestinal mucosa in vivo, which correlated with altered fatty acid metabolism. Our study links the HIV/SIV infection–induced fatty acid enzyme ACSS2 to HIV latency and identifies histone lysine crotonylation as a novel epigenetic regulator for HIV transcription that can be targeted for HIV eradication.

Authors

Guochun Jiang, Don Nguyen, Nancie M. Archin, Steven A. Yukl, Gema Méndez-Lagares, Yuyang Tang, Maher M. Elsheikh, George R. Thompson III, Dennis J. Hartigan-O’Connor, David M. Margolis, Joseph K. Wong, Satya Dandekar

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Figure 2

Histone crotonylation by ACSS2 induction reprograms histone tails at the HIV LTR, and suppression of ACSS2 dampens reactivation of latent HIV.

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Histone crotonylation by ACSS2 induction reprograms histone tails at the...
(A) Induction of ACSS2 increased global expression of H3K4 crotonylation (H3K4Cr), H3K4 acetylation (H3K4Ac), and H3K18 acetylation (H3K18Ac) while decreasing expression of H3K27 methylation (H3K27Me) after Na-Cro treatment for 4 hours in J-Lat A1 cells. Immunoblots are representative of 3 independent experiments. (B) Protein expression after Na-Cro treatment for 4 hours in J-Lat A1 cells. Immunoblots are representative of 3 independent experiments. (C–E) Na-Cro addition induced histone crotonylation at H3K4 (C) and histone acetylation at H3K4 (E) while suppressing histone trimethylation at H3K27 (D) at the HIV LTR. ChIP assay was performed using anti-H3K4Cr (n = 3), anti-H3K4Ac (n = 3), and anti-H3K27Me3 (n = 6) antibodies after J-Lat A1 cells were treated with 30 mM Na-Cro for 4 hours. PCR primers were specific for the HIV LTR region. **P < 0.01 versus control treatment. (F) Reactivation of latent HIV was examined using different epigenetic modifiers. J-Lat A1 cells were treated with 30 mM Na-Cro, 500 nM vorinostat, 2 μM GSK343, or 5 μM AZA-dC. HIV expression was measured by RT-qPCR. **P < 0.01 and ***P < 0.001 versus control treatment (n = 3). (G) Pretreatment with the ACSS2 inhibitor AR12 (5 μM for 30 minutes) dampened histone crotonylation–induced reactivation of latent HIV. ***P < 0.001 versus control treatment; ###P < 0.001 versus Na-Cro treatment (n = 4). (H) Suppression of ACSS2 by AR12 decreased histone crotonylation at the HIV LTR. J-Lat A1 cells were pretreated with 5 μM AR12 for 30 minutes, followed by Na-Cro (20 mM) addition and incubation for 4 hours. Crotonylation of H3K18Cr at the HIV LTR was measured by ChIP assay. **P < 0.01 versus control treatment; ##P < 0.01 versus Na-Cro treatment (n = 3). The data were analyzed with a Student’s t test or 1-way ANOVA.