Transcriptional behavior of the HIV-1 promoter in context of the BACH2 prominent proviral integration gene

Abstract VPAC1 is a 7-transmembrane G-protein-coupled neuroendocrine receptor previously shown to transduce a facilitation signal for HIV-1 infection (Branch DR, et al., AIDS.2002;16:309–319). VPAC2, a related receptor, has been reported to have opposing function when compared to VPAC1 (Xia M, et al., J Immunol.1996;157:1132–1138; Tsutsumi M, et al., Diabetes.2002;51:1453–1460). We therefore examined whether stimulation of VPAC2, in contrast to VPAC1, may act to inhibit HIV-1 infection. Using three different and specific agonists of VPAC2, helodermin, RO 25-1553, and R3P55, daily treatment with low concentrations (10−9M) resulted in ~75% to 95% inhibition of either X4 or R5 HIV-1 productive infection in cell lines or primary peripheral blood mononuclear cells. The agonists VIP, PACAP, and secretin, that stimulate the two other VPAC receptor family members VPAC1 and PAC1, did not inhibit HIV-1 infection. Also, Hut78 cells, that lack VPAC2 and are infected by HIV-1, show no effect of VPAC2 agonists on HIV-1 infection. However, Hut78 cells transfected with human VPAC2 cDNA to overexpress this receptor become resistant to HIV-1 infection when treated with VPAC2 agonists. VPAC2-mediated inhibition of productive HIV-1 infection was not due to effects on CD4 or chemokine co-receptor expression, cell growth, or apoptosis. Treatment with VPAC2 agonists also did not inhibit HIV-1 entry into the host cell. However, compared to untreated or cells treated with VPAC1-specific agonists, VPAC2 stimulation profoundly inhibits HIV-1 proviral DNA integration into the host genome. The block in integration was found to be due to the ability of agonists to VPAC2 to inhibit formation of 2-LTR circles, required for HIV-1 integration within the nucleus. We conclude that VPAC2-specific agonists are strongly inhibitory for productive HIV-1 infection. Furthermore, this inhibition is mediated by suppression of 2-LTR circle formation preventing proviral integration. Agonists of VPAC2 appear to be excellent candidates for future development as possible drugs for the amelioration of treatments aimed at the prevention of HIV/AIDS.

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HIV-1 Latency Due to the Site of Proviral Integration

Virology ◽

10.1006/viro.1993.1545 ◽

1993 ◽

Vol 196 (2) ◽

pp. 849-854 ◽

Cited By ~ 45

Author(s):

Barbara J. Winslow ◽

Roger J. Pomerantz ◽

Omar Bagasra ◽

Didier Trono

Keyword(s):

Proviral Integration ◽

Hiv 1

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Loss of the Brm-Type SWI/SNF Chromatin Remodeling Complex Is a Strong Barrier to the Tat-Independent Transcriptional Elongation of Human Immunodeficiency Virus Type 1 Transcripts

Journal of Virology ◽

10.1128/jvi.00742-09 ◽

2009 ◽

Vol 83 (22) ◽

pp. 11569-11580 ◽

Cited By ~ 17

Author(s):

Taketoshi Mizutani ◽

Aya Ishizaka ◽

Mariko Tomizawa ◽

Takuya Okazaki ◽

Nobutake Yamamichi ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Cell Lines ◽

Competent Cell ◽

Transcriptional Elongation ◽

Gfp Expression ◽

Proviral Integration ◽

Hiv Transcription ◽

Immunodeficiency Virus ◽

Hiv 1

ABSTRACT To elucidate the epigenetic regulation of Tat-independent human immunodeficiency virus (HIV) transcription following proviral integration, we constructed an HIV type 1 (HIV-1)-based replication-defective viral vector that expresses a reporter green fluorescent protein (GFP) product from its intact long terminal repeat (LTR). We transduced this construct into human tumor cell lines that were either deficient in or competent for the Brm-type SWI/SNF complex. One day after transduction, single cells that expressed GFP were sorted, and the GFP expression profiles originating from each of these clones were analyzed. Unlike clones of the SWI/SNF-competent cell line, which exhibited clear unimodal expression patterns in all cases, many clones originating from Brm-deficient cell lines either showed a broad-range distribution of GFP expression or were fully silenced. The resorting of GFP-negative populations of these isolated clones showed that GFP silencing is either reversible or irreversible depending upon the proviral integration sites. We further observed that even in these silenced clones, proviral gene transcription initiates to accumulate short transcripts of around 60 bases in length, but no elongation occurs. We found that this termination is caused by tightly closed nucleosome-1 (nuc-1) at the 5′ LTR. Also, nuc-1 is remodeled by exogenous Brm in some integrants. From these results, we propose that Brm is required for the occasional transcriptional elongation of the HIV-1 provirus in the absence of Tat. Since the Brm-type SWI/SNF complex is expressed at marginal levels in resting CD4+ T cells and is drastically induced upon CD4+ T-cell activation, we speculate that it plays crucial roles in the early Tat-independent phase of HIV transcription in affected patients.

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HIV-1 replication is controlled at the level of T cell activation and proviral integration.

The EMBO Journal ◽

10.1002/j.1460-2075.1990.tb08274.x ◽

1990 ◽

Vol 9 (5) ◽

pp. 1551-1560 ◽

Cited By ~ 530

Author(s):

M. Stevenson ◽

T.L. Stanwick ◽

M.P. Dempsey ◽

C.A. Lamonica

Keyword(s):

T Cell ◽

T Cell Activation ◽

Cell Activation ◽

Proviral Integration ◽

Hiv 1

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Analysis of HIV quasispecies and virological outcome of an HIV D+/R+ kidney–liver transplantation

Virology Journal ◽

10.1186/s12985-021-01730-w ◽

2022 ◽

Vol 19 (1) ◽

Author(s):

Gabriella Rozera ◽

Ubaldo Visco-Comandini ◽

Emanuela Giombini ◽

Francesco Santini ◽

Federica Forbici ◽

...

Keyword(s):

Liver Transplantation ◽

Drug Resistance ◽

Immunosuppressive Treatment ◽

Proviral Integration ◽

Infected Woman ◽

Cell Counts ◽

Integration Sites ◽

Kidney Liver ◽

Hiv 1

Abstract Introduction Transplantation among HIV positive patients may be a valuable therapeutic intervention. This study involves an HIV D+/R+ kidney–liver transplantation, where PBMC-associated HIV quasispecies were analyzed in donor and transplant recipients (TR) prior to transplantation and thereafter, together with standard viral monitoring. Methods The donor was a 54 year of age HIV infected woman: kidney and liver recipients were two HIV infected men, aged 49 and 61. HIV quasispecies in PBMC was analyzed by ultra-deep sequencing of V3 env region. During TR follow-up, plasma HIV-1 RNA, HIV-1 DNA in PBMC, analysis of proviral integration sites and drug-resistance genotyping were performed. Other virological and immunological monitoring included CMV and EBV DNA quantification in blood and CD4 T cell counts. Results Donor and TR were all ART-HIV suppressed at transplantation. Thereafter, TR maintained a nearly suppressed HIV-1 viremia, but HIV-1 RNA blips and the increase of proviral integration sites in PBMC attested some residual HIV replication. A transient peak in HIV-1 DNA occurred in the liver recipient. No major changes of drug-resistance genotype were detected after transplantation. CMV and EBV transient reactivations were observed only in the kidney recipient, but did not require specific treatment. CD4 counts remained stable. No intermixed quasispecies between donor and TR was observed at transplantation or thereafter. Despite signs of viral evolution in TR, HIV genetic heterogeneity did not increase over the course of the months of follow up. Conclusions No evidence of HIV superinfection was observed in the donor nor in the recipients. The immunosuppressive treatment administrated to TR did not result in clinical relevant viral reactivations.

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CRISPR-Cas9-based Genome Engineering to Generate Jurkat Reporter Models for HIV-1 Infection with Selected Proviral Integration Sites

Journal of Visualized Experiments ◽

10.3791/58572 ◽

2018 ◽

Cited By ~ 2

Author(s):

Julia K. Bialek ◽

Thomas Walther ◽

Joachim Hauber ◽

Ulrike C. Lange

Keyword(s):

Genome Engineering ◽

Proviral Integration ◽

Integration Sites ◽

Hiv 1

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Genomic Sites of Human Immunodeficiency Virus Type 2 (HIV-2) Integration: Similarities to HIV-1 In Vitro and Possible Differences In Vivo

Journal of Virology ◽

10.1128/jvi.00604-06 ◽

2006 ◽

Vol 80 (15) ◽

pp. 7316-7321 ◽

Cited By ~ 25

Author(s):

Adam MacNeil ◽

Jean-Louis Sankalé ◽

Seema Thakore Meloni ◽

Abdoulaye Dieng Sarr ◽

Souleymane Mboup ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Site Selection ◽

Integration Site ◽

Proviral Integration ◽

Immunodeficiency Virus ◽

Hiv 1 ◽

Integration Site Selection

ABSTRACT Retroviruses have distinct preferences in integration site selection in the host cell genome during in vitro infection, with human immunodeficiency virus type 1 (HIV-1) integration strongly favoring transcriptional units. Additionally, studies with HIV-1 have shown that the genomic site of proviral integration may impact viral replication, with integration in heterochromatin associated with a block in viral transcription. HIV-2 is less pathogenic than HIV-1 and is believed to have a lower replication rate in vivo. Although differences in integration site selection between HIV-2 and HIV-1 could potentially explain the attenuated pathogenicity of HIV-2, no studies have characterized integration site selection by HIV-2. In this study, we mapped 202 HIV-2 integration sites during in vitro infection of peripheral blood mononuclear cells with a primary HIV-2 isolate. In addition, we assayed for in vivo proviral integration within heterochromatin in 21 HIV-1-infected subjects and 23 HIV-2-infected subjects, using an alphoid repeat PCR assay. During in vitro infection, HIV-2 displayed integration site preferences similar to those previously reported for HIV-1. Notably, 82% of HIV-2 integrations mapped to Refseq genes, and integration strongly favored regions of the genome with high gene density and high GC content. Though rare, the proportion of HIV-2 subjects with evidence of proviral integration within heterochromatin in vivo was higher than that of HIV-1-infected subjects. It is therefore possible that integration site selection may play a role in the differences in HIV-1 and HIV-2 in vivo pathogenesis.

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