TRIM28 promotes HIV-1 latency by SUMOylating CDK9 and inhibiting P-TEFb

  1. Xiancai Ma
  2. Tao Yang
  3. Yuewen Luo
  4. Liyang Wu
  5. Yawen Jiang
  6. Zheng Song
  7. Ting Pan
  8. Bingfeng Liu
  9. Guangyan Liu
  10. Jun Liu
  11. Fei Yu
  12. Zhangping He
  13. Wanying Zhang
  14. Jinyu Yang
  15. Liting Liang
  16. Yuanjun Guan
  17. Xu Zhang
  18. Linghua Li
  19. Weiping Cai
  20. Xiaoping Tang
  21. Song Gao
  22. Kai Deng
  23. Hui Zhang  Is a corresponding author
  1. Sun Yat-sen University, China
  2. Shenyang Medical College, China
  3. Sun Yat-Sen University Cancer Center, China
  4. Guangzhou 8th People's Hospital, China

Abstract

Comprehensively elucidating the molecular mechanisms of human immunodeficiency virus type 1 (HIV-1) latency is a priority to achieve a functional cure. As current 'shock' agents failed to efficiently reactivate the latent reservoir, it is important to discover new targets for developing more efficient latency-reversing agents (LRAs). Here we found that TRIM28 potently suppresses HIV-1 expression by utilizing both SUMO E3 ligase activity and epigenetic adaptor function. Through global site-specific SUMO-MS study and serial SUMOylation assays, we identified that P-TEFb catalytic subunit CDK9 is significantly SUMOylated by TRIM28 with SUMO4. The Lys44, Lys56 and Lys68 residues on CDK9 are SUMOylated by TRIM28, which inhibits CDK9 kinase activity or prevents P-TEFb assembly by directly blocking the interaction between CDK9 and Cyclin T1, subsequently inhibits viral transcription and contributes to HIV-1 latency. The manipulation of TRIM28 and its consequent SUMOylation pathway could be the target for developing LRAs.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files.

Article and author information

Author details

  1. Xiancai Ma

    Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  2. Tao Yang

    Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  3. Yuewen Luo

    Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  4. Liyang Wu

    Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  5. Yawen Jiang

    Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  6. Zheng Song

    Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  7. Ting Pan

    Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  8. Bingfeng Liu

    Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  9. Guangyan Liu

    College of Basic Medical Sciences, Shenyang Medical College, Shenyang, China
    Competing interests
    The authors declare that no competing interests exist.
  10. Jun Liu

    Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  11. Fei Yu

    Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  12. Zhangping He

    Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  13. Wanying Zhang

    Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  14. Jinyu Yang

    State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  15. Liting Liang

    Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  16. Yuanjun Guan

    Core Laboratory Platform for Medical Science, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  17. Xu Zhang

    Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  18. Linghua Li

    Department of Infectious Diseases, Guangzhou 8th People's Hospital, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  19. Weiping Cai

    Department of Infectious Diseases, Guangzhou 8th People's Hospital, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  20. Xiaoping Tang

    Department of Infectious Diseases, Guangzhou 8th People's Hospital, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  21. Song Gao

    State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7427-6681
  22. Kai Deng

    Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  23. Hui Zhang

    Institute of Human Virology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    For correspondence
    zhangh92@mail.sysu.edu.cn
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3620-610X

Funding

National Special Research Program of China for Important Infectious Diseases (2017ZX10202102)

  • Hui Zhang

Important Key Program of Natural Science Foundation of China (81730060)

  • Hui Zhang

International Collaboration Program of Natural Science Foundation of China and US NIH (81561128007)

  • Hui Zhang

Joint-innovation Program in Healthcare for Special Scientific Research Projects of Guangzhou (201803040002)

  • Hui Zhang

National Special Research Program of China for Important Infectious Diseases (2018ZX10302103)

  • Hui Zhang

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Human subjects: Chronically HIV-1-infected participants sampled by this study were recruited from Department of Infectious Diseases in Guangzhou 8th People's Hospital, Guangzhou. The Ethics Review Board of Sun Yat-Sen University and the Ethics Review Board of Guangzhou 8th People's Hospital approved this study. All the participants were given written informed consent with approval of the Ethics Committees. The enrollment of HIV-1-infected individuals was based on the criteria of prolonged suppression of plasma HIV-1 viremia on cART, which is undetectable plasma HIV-1 RNA levels (less than 50 copies/ml) for a minimum of six months, and having high CD4+ T cell count (at least 350 cells/mm3). Blood samples from healthy individuals were obtained from Guangzhou Blood Center. We did not have any interaction with the healthy individuals or protected information, and therefore no informed consent was required. The statement was also included in the Materials and Methods section.

Copyright

© 2019, Ma et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 4,243
    views
  • 869
    downloads
  • 83
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Xiancai Ma
  2. Tao Yang
  3. Yuewen Luo
  4. Liyang Wu
  5. Yawen Jiang
  6. Zheng Song
  7. Ting Pan
  8. Bingfeng Liu
  9. Guangyan Liu
  10. Jun Liu
  11. Fei Yu
  12. Zhangping He
  13. Wanying Zhang
  14. Jinyu Yang
  15. Liting Liang
  16. Yuanjun Guan
  17. Xu Zhang
  18. Linghua Li
  19. Weiping Cai
  20. Xiaoping Tang
  21. Song Gao
  22. Kai Deng
  23. Hui Zhang
(2019)
TRIM28 promotes HIV-1 latency by SUMOylating CDK9 and inhibiting P-TEFb
eLife 8:e42426.
https://doi.org/10.7554/eLife.42426

Share this article

https://doi.org/10.7554/eLife.42426

Further reading

    1. Genetics and Genomics
    2. Microbiology and Infectious Disease
    Iti Mehta, Jacob B Hogins ... Larry Reitzer
    Research Article

    Polyamines are biologically ubiquitous cations that bind to nucleic acids, ribosomes, and phospholipids and, thereby, modulate numerous processes, including surface motility in Escherichia coli. We characterized the metabolic pathways that contribute to polyamine-dependent control of surface motility in the commonly used strain W3110 and the transcriptome of a mutant lacking a putrescine synthetic pathway that was required for surface motility. Genetic analysis showed that surface motility required type 1 pili, the simultaneous presence of two independent putrescine anabolic pathways, and modulation by putrescine transport and catabolism. An immunological assay for FimA—the major pili subunit, reverse transcription quantitative PCR of fimA, and transmission electron microscopy confirmed that pili synthesis required putrescine. Comparative RNAseq analysis of a wild type and ΔspeB mutant which exhibits impaired pili synthesis showed that the latter had fewer transcripts for pili structural genes and for fimB which codes for the phase variation recombinase that orients the fim operon promoter in the ON phase, although loss of speB did not affect the promoter orientation. Results from the RNAseq analysis also suggested (a) changes in transcripts for several transcription factor genes that affect fim operon expression, (b) compensatory mechanisms for low putrescine which implies a putrescine homeostatic network, and (c) decreased transcripts of genes for oxidative energy metabolism and iron transport which a previous genetic analysis suggests may be sufficient to account for the pili defect in putrescine synthesis mutants. We conclude that pili synthesis requires putrescine and putrescine concentration is controlled by a complex homeostatic network that includes the genes of oxidative energy metabolism.

    1. Biochemistry and Chemical Biology
    2. Microbiology and Infectious Disease
    Eva Herdering, Tristan Reif-Trauttmansdorff ... Ruth Anne Schmitz
    Research Article

    Glutamine synthetases (GS) are central enzymes essential for the nitrogen metabolism across all domains of life. Consequently, they have been extensively studied for more than half a century. Based on the ATP-dependent ammonium assimilation generating glutamine, GS expression and activity are strictly regulated in all organisms. In the methanogenic archaeon Methanosarcina mazei, it has been shown that the metabolite 2-oxoglutarate (2-OG) directly induces the GS activity. Besides, modulation of the activity by interaction with small proteins (GlnK1 and sP26) has been reported. Here, we show that the strong activation of M. mazei GS (GlnA1) by 2-OG is based on the 2-OG dependent dodecamer assembly of GlnA1 by using mass photometry (MP) and single particle cryo-electron microscopy (cryo-EM) analysis of purified strep-tagged GlnA1. The dodecamer assembly from dimers occurred without any detectable intermediate oligomeric state and was not affected in the presence of GlnK1. The 2.39 Å cryo-EM structure of the dodecameric complex in the presence of 12.5 mM 2-OG demonstrated that 2-OG is binding between two monomers. Thereby, 2-OG appears to induce the dodecameric assembly in a cooperative way. Furthermore, the active site is primed by an allosteric interaction cascade caused by 2-OG-binding towards an adaption of an open active state conformation. In the presence of additional glutamine, strong feedback inhibition of GS activity was observed. Since glutamine dependent disassembly of the dodecamer was excluded by MP, feedback inhibition most likely relies on the binding of glutamine to the catalytic site. Based on our findings, we propose that under nitrogen limitation the induction of M. mazei GS into a catalytically active dodecamer is not affected by GlnK1 and crucially depends on the presence of 2-OG.