CMPK2 and BCL-G are associated with type 1 interferon–induced HIV restriction in humans

El-Diwany, Ramy; Soliman, Mary; Sugawara, Sho K.; Breitwieser, Florian P.; Skaist, Alyza; Coggiano, Candelaria; Sangal, Neel; Chattergoon, Michael A.; Bailey, Justin R.; Siliciano, Robert F.; Blankson, Joel N.; Ray, Stuart C.; Wheelan, Sarah J.; Thomas, David L.; Balagopal, Ashwin

doi:10.1126/sciadv.aat0843

Cited by 65 publications

(87 citation statements)

References 45 publications

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“…Of note, TRAPPC4 was shown to regulate activity and nuclear localisation of ERK1/2 in human colorectal cancer cells 36 , while Bcl-G itself is known to restrict LPS-induced ERK1/2 signalling in mouse macrophages 37 . Another example of an immunoregulatory function of BCL-G comes from a 2018 study, which identified BCL-G as a novel restriction factor for HIV infection in humans during the course IFN-α2b therapy 38 . Most recently, Nguyen and colleagues reported that loss of Bcl-G aggravated development of colitisassociated cancer in mice, likely through disruption of tissue-protective functions of IEC as intratumoural infiltration by CD45 + leucocytes was comparable between wild-type and Bcl-G −/− hosts 6 .…”

Section: Discussionmentioning

confidence: 99%

Human BCL-G regulates secretion of inflammatory chemokines but is dispensable for induction of apoptosis by IFN-γ and TNF-α in intestinal epithelial cells

et al. 2020

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Proteins of the BCL-2 family are evolutionarily conserved modulators of apoptosis that function as sensors of cellular integrity. Over the past three decades multiple BCL-2 family members have been identified, many of which are now fully incorporated into regulatory networks governing the mitochondrial apoptotic pathway. For some, however, an exact role in cell death signalling remains unclear. One such 'orphan' BCL-2 family member is BCL-G (or BCL2L14). In this study we analysed gastrointestinal expression of human BCL-G in health and disease states, and investigated its contribution to inflammation-induced tissue damage by exposing intestinal epithelial cells (IEC) to IFN-γ and TNF-α, two pro-inflammatory mediators associated with gut immunopathology. We found that both BCL-G splice variants -BCL-G S (short) and BCL-G L (long)were highly expressed in healthy gut tissue, and that their mRNA levels decreased in active inflammatory bowel diseases (for BCL-G S ) and colorectal cancer (for BCL-G S/L ). In vitro studies revealed that IFN-γ and TNF-α synergised to upregulate BCL-G S/L and to trigger apoptosis in colonic epithelial cell lines and primary human colonic organoids. Using RNAi, we showed that synergistic induction of IEC death was STAT1-dependent while optimal expression of BCL-G S/L required STAT1, NF-κB/p65 and SWI/SNF-associated chromatin remodellers BRM and BRG1. To test the direct contribution of BCL-G to the effects of IFN-γ and TNF-α on epithelial cells, we used RNAi-and CRISPR/Cas9-based perturbations in parallel with isoform-specific overexpression of BCL-G, and found that BCL-G was dispensable for Th1 cytokine-induced apoptosis of human IEC. Instead, we discovered that depletion of BCL-G differentially affected secretion of inflammatory chemokines CCL5 and CCL20, thus uncovering a non-apoptotic immunoregulatory function of this BCL-2 family member. Taken together, our data indicate that BCL-G may be involved in shaping immune responses in the human gut in health and disease states through regulation of chemokine secretion rather than intestinal apoptosis.

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Section: Discussionmentioning

confidence: 99%

Human BCL-G regulates secretion of inflammatory chemokines but is dispensable for induction of apoptosis by IFN-γ and TNF-α in intestinal epithelial cells

et al. 2020

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“…Figure 3B). In addition, a list of the virus-specific antiviral-related genes was identified, including PARP10 [21] and CMPK2 [22] for SARS-CoV-2, BST2 [23], ITITM1 and USP41 for SARS, and PARP4 [24] for MERS-CoV ( Figure 3B). Secondly, we further used a set of 113 human cytokines to quantify host inflammation responses between three viruses.…”

Section: Mers-covmentioning

confidence: 99%

Comparative transcriptome analysis reveals the intensive early-stage responses of host cells to SARS-CoV-2 infection

Sun

et al. 2020

Preprint

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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a widespread outbreak of highly pathogenic COVID-19. It is therefore important and timely to characterize interactions between the virus and host cell at the molecular level to understand its disease pathogenesis. To gain insights, we performed high-throughput sequencing that generated time-series data simultaneously for bioinformatics analysis of virus genomes and host transcriptomes implicated in SARS-CoV-2 infection. Our analysis results showed that the rapid growth of the virus was accompanied by an early intensive response of host genes. We also systematically compared the molecular footprints of the host cells in response to SARS-CoV-2, SARS-CoV and MERS-CoV. Upon infection, SARS-CoV-2 induced hundreds of up-regulated host genes hallmarked by a significant cytokine production followed by virus-specific host antiviral responses. While the cytokine and antiviral responses triggered by SARS-CoV and MERS-CoV were only observed during the late stage of infection, the host antiviral responses during the SARS-CoV-2 infection were gradually enhanced lagging behind the production of cytokine. The early rapid host responses were potentially attributed to the high efficiency of SARS-CoV-2 entry into host cells, underscored by evidence of a remarkably up-regulated gene expression of TPRMSS2 soon after infection. Taken together, our findings provide novel molecular insights into the mechanisms underlying the infectivity and pathogenicity of SARS-CoV-2.

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“…However, the enhanced resistance to interferon/higher infectivity titer exhibited by the lab adapted stains was higher than that of TF viruses 19 . IFN-α upregulates the expression of the several well-known HIV-1 and SIV restriction factors such as TRIM5α (tripartite motif-containing protein 5α), APOBEC3G (apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like 3G), tetherin and SAMHD1 (SAM and HD domain-containing protein 1) in addition to HIV-1 resistance factors such as myxovirus resistance 2 (MX2), schlafen 11 (SLFN11), IFN-induced transmembrane (IFITM) proteins 38 , BCL-G 40 and Cytidine/Uridine Monophosphate Kinase 2 (CMPK2) 40,41 , which constrain HIV-1 replication at different steps. Several studies have reported that early release of IFN-α suppresses the viral load during the acute phase of HIV-1 infection.…”

Section: Discussionmentioning

confidence: 99%

In vitro replicative fitness of early Transmitted founder HIV-1 variants and sensitivity to Interferon alpha

Ashokkumar

Sonawane

Sperk

et al. 2020

Sci Rep

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Type I interferons, particularly interferon-alpha (IFN-α), play a vital role in the host's anti-viral defensesby interfering with viral replication. However, the virus rapidly evolves to exploit the IFN-α response for its replication, spread, and pathogenic function. In this study, we attempted to determine IFN-α susceptibility and productivity of infectious transmitted/founder (TF) (n = 8) and non-transmitted (NT) viruses (n = 8) derived from HIV-1 infected infants. Independent experiments were carried out to determine IFN-α resistance, replication fitness, and viral productivity in CD4 + T cells over a short period. In vitro studies showed that TF viruses were resistant to IFN-α during the very near moment of transmission, but in the subsequent time points, they became susceptible to IFN-α. We did not observe much difference in replicative fitness of the TF viruses in cultures treated with and without IFN-α, but the difference was significant in the case of NT viruses obtained from the same individual. Despite increased susceptibility to IFN-α, NT viruses produced more viral particles than TF viruses. Similar results were also obtained in cultures treated with maraviroc (MVC). The study identified unique characteristics of TF viruses thus prompting further investigation into virus-host interaction occurring during the early stages of HIV infection. Determination of saturation point for maraviroc. TZM-bl and GHOST (3) CCR5+ (Hi-5) celllines were employed to assess the sensitivity of chimeric viruses to the CCR5 antagonist, maraviroc. Briefly, 10 4 TZM-bl and Hi-5 cells were plated in each well of a 96-well plate 24 hours before infection. Cells were incubated with different dilutions of MVC (10-fold dilutions starting from 10 μM up to 10 −3 nM) overnight at 37 °C before inoculation with chimeric viruses. Virus infected cells cultured in the absence of MVC served as negative control. At 48 hours post-infection, cells were lysed with 1X passive lysis buffer (Bright-Glo luciferase assay system) and luciferase expression was measured using an Infinite M200Pro plate reader (Tecan, Switzerland) as the number of relative light units (RLU). All conditions were tested in triplicate in each of at least two independent experiments.Single round virus replication kinetics assay. For single cycle infectivity assay, virus stocks normalized by TCID 50 and further diluted to 0.01 MOI were used to infect TZM-bl cells (1 × 10 4 cells/well) in triplicates in the presence of DEAE dextran (10 μg/mL). Cells were pre-incubated for 2 hours with different concentrations (0, 250, 500 and 750 IU/mL) of IFN-α at 37 °C before infection with chimeric viruses. 48 hours after infection, luciferase activity was measured using the Bright-Glo luciferase assay system (Promega). Multiple round virus replication kinetics assay. CD4 + T cells were isolated from buffy coats by deplet-ing CD8 + T cells and stimulated with PHA (20 μg/mL) in the presence of 20 U/mL IL-2 in RPMI medium containing 10% FBS at a concentration of 1 × 10 6 cells per milliliter....

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CMPK2 and BCL-G are associated with type 1 interferon–induced HIV restriction in humans

Abstract: We identified two genes induced by type 1 interferon in activated CD4+ T cells that are associated with HIV restriction in humans.

Cited by 65 publications

References 45 publications

Human BCL-G regulates secretion of inflammatory chemokines but is dispensable for induction of apoptosis by IFN-γ and TNF-α in intestinal epithelial cells

Human BCL-G regulates secretion of inflammatory chemokines but is dispensable for induction of apoptosis by IFN-γ and TNF-α in intestinal epithelial cells

Comparative transcriptome analysis reveals the intensive early-stage responses of host cells to SARS-CoV-2 infection

In vitro replicative fitness of early Transmitted founder HIV-1 variants and sensitivity to Interferon alpha

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