SAMHD1 Impairs HIV-1 Gene Expression and Negatively Modulates Reactivation of Viral Latency in CD4 + T Cells

Jenna M Antonucci; Sun Hee Kim; Corine St Gelais; Serena Bonifati; Tai-Wei Li; Olga Buzovetsky; Kirsten M Knecht; Alice A Duchon; Yong Xiong; Karin Musier-Forsyth; Li Wu

doi:10.1128/jvi.00292-18

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SAMHD1 Impairs HIV-1 Gene Expression and Negatively Modulates Reactivation of Viral Latency in CD4 + T Cells

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SAMHD1 Impairs HIV-1 Gene Expression and Negatively Modulates Reactivation of Viral Latency in CD4 + T Cells

Jenna M Antonucci, Sun Hee Kim, Corine St Gelais, Serena Bonifati, Tai-Wei Li, Olga Buzovetsky, Kirsten M Knecht, Alice A Duchon, Yong Xiong, Karin Musier-Forsyth, …

Journal of virology, Vol.92(15), e00292-18

08/01/2018

DOI: 10.1128/jvi.00292-18

PMCID: PMC6052313

PMID: 29793958

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Abstract

Sterile alpha motif and HD domain-containing protein 1 (SAMHD1) restricts human immunodeficiency virus type 1 (HIV-1) replication in nondividing cells by degrading intracellular deoxynucleoside triphosphates (dNTPs). SAMHD1 is highly expressed in resting CD4 T cells, which are important for the HIV-1 reservoir and viral latency; however, whether SAMHD1 affects HIV-1 latency is unknown. Recombinant SAMHD1 binds HIV-1 DNA or RNA fragments , but the function of this binding remains unclear. Here we investigate the effect of SAMHD1 on HIV-1 gene expression and reactivation of viral latency. We found that endogenous SAMHD1 impaired HIV-1 long terminal repeat (LTR) activity in monocytic THP-1 cells and HIV-1 reactivation in latently infected primary CD4 T cells. Overexpression of wild-type (WT) SAMHD1 suppressed HIV-1 LTR-driven gene expression at a transcriptional level. Tat coexpression abrogated SAMHD1-mediated suppression of HIV-1 LTR-driven luciferase expression. SAMHD1 overexpression also suppressed the LTR activity of human T-cell leukemia virus type 1 (HTLV-1), but not that of murine leukemia virus (MLV), suggesting specific suppression of retroviral LTR-driven gene expression. WT SAMHD1 bound to proviral DNA and impaired reactivation of HIV-1 gene expression in latently infected J-Lat cells. In contrast, a nonphosphorylated mutant (T592A) and a dNTP triphosphohydrolase (dNTPase) inactive mutant (H206D R207N [HD/RN]) of SAMHD1 failed to efficiently suppress HIV-1 LTR-driven gene expression and reactivation of latent virus. Purified recombinant WT SAMHD1, but not the T592A and HD/RN mutants, bound to fragments of the HIV-1 LTR These findings suggest that SAMHD1-mediated suppression of HIV-1 LTR-driven gene expression potentially regulates viral latency in CD4 T cells. A critical barrier to developing a cure for HIV-1 infection is the long-lived viral reservoir that exists in resting CD4 T cells, the main targets of HIV-1. The viral reservoir is maintained through a variety of mechanisms, including regulation of the HIV-1 LTR promoter. The host protein SAMHD1 restricts HIV-1 replication in nondividing cells, but its role in HIV-1 latency remains unknown. Here we report a new function of SAMHD1 in regulating HIV-1 latency. We found that SAMHD1 suppressed HIV-1 LTR promoter-driven gene expression and reactivation of viral latency in cell lines and primary CD4 T cells. Furthermore, SAMHD1 bound to the HIV-1 LTR and in a latently infected CD4 T-cell line, suggesting that the binding may negatively modulate reactivation of HIV-1 latency. Our findings indicate a novel role for SAMHD1 in regulating HIV-1 latency, which enhances our understanding of the mechanisms regulating proviral gene expression in CD4 T cells.

Gene Expression Regulation, Viral - physiology

HIV Long Terminal Repeat - physiology

SAM Domain and HD Domain-Containing Protein 1 - metabolism

Jurkat Cells

Humans

CD4-Positive T-Lymphocytes - metabolism

THP-1 Cells

Mutation, Missense

Virus Latency - physiology

Transcription, Genetic - physiology

HIV-1 - physiology

HEK293 Cells

CD4-Positive T-Lymphocytes - virology

SAM Domain and HD Domain-Containing Protein 1 - genetics

Amino Acid Substitution

Details

Title: Subtitle: SAMHD1 Impairs HIV-1 Gene Expression and Negatively Modulates Reactivation of Viral Latency in CD4 + T Cells
Creators: Jenna M Antonucci - Department of Microbiology, The Ohio State University, Columbus, Ohio, USA
Sun Hee Kim - Department of Veterinary Biosciences, The Ohio State University, Columbus, Ohio, USA
Corine St Gelais - Department of Veterinary Biosciences, The Ohio State University, Columbus, Ohio, USA
Serena Bonifati - Department of Veterinary Biosciences, The Ohio State University, Columbus, Ohio, USA
Tai-Wei Li - Department of Veterinary Biosciences, The Ohio State University, Columbus, Ohio, USA
Olga Buzovetsky - Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut, USA
Kirsten M Knecht - Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut, USA
Alice A Duchon - Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio, USA
Yong Xiong - Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut, USA
Karin Musier-Forsyth - Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio, USA
Li Wu - Center for RNA Biology, The Ohio State University, Columbus, Ohio, USA
Resource Type: Journal article
Publication Details: Journal of virology, Vol.92(15), e00292-18
Publisher: United States
DOI: 10.1128/jvi.00292-18
PMID: 29793958
PMCID: PMC6052313
ISSN: 1098-5514
eISSN: 1098-5514
Grant note: R01 GM113887 / NIGMS NIH HHS F31 GM119178 / NIGMS NIH HHS T32 GM008283 / NIGMS NIH HHS R01 GM128212 / NIGMS NIH HHS R01 AI104483 / NIAID NIH HHS T32 GM007223 / NIGMS NIH HHS R01 AI120209 / NIAID NIH HHS
Language: English
Date published: 08/01/2018
Academic Unit: Microbiology and Immunology
Record Identifier: 9984002386702771

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