Covalent modification by SUMO is required for efficient disruption of PML oncogenic domains by Kaposi's sarcoma-associated herpesvirus latent protein LANA2

Marcos-Villar, Laura; Campagna, Michela; Lopitz‐Otsoa, Fernando; Gallego, Pedro Pablo Ferrer; González-Santamaría, José; González, Dolores; Rodríguez, Manuel Sánchez; Rivas, Carmen

doi:10.1099/vir.0.024984-0

Cited by 32 publications

(30 citation statements)

References 38 publications

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“…Specifically, expression of vIRF3 increases the levels of SUMO2 ubiquitination-modified promyelocytic leukemia (PML) gene product oncogenic domains (PODs) (31). Furthermore, vIRF3 is covalently conjugated to SUMO1 and SUMO2 in vitro and in KSHV latently infected B cells (32). vIRF3 is also capable of inhibiting SUMOylation of the pRb proteins pRb, p107, and p130 (33).…”

Section: Discussionmentioning

confidence: 99%

Kaposi's Sarcoma-Associated Herpesvirus Viral Interferon Regulatory Factor 1 Interacts with a Member of the Interferon-Stimulated Gene 15 Pathway

Jacobs

Stopford

West

et al. 2015

J Virol

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Kaposi's sarcoma-associated herpesvirus (KSHV) is a gammaherpesvirus known to establish lifelong latency in the human host. We and others have previously shown that three KSHV homologs of cellular interferon regulatory factors (IRFs), known as viral IRFs (vIRFs), participate in evasion of the host interferon (IFN) response. We report that vIRF1 interacts with the cellular interferon-stimulated gene 15 (ISG15) E3 ligase, HERC5, in the context of Toll-like receptor 3 (TLR3) activation and IFN induction. The ISG15 protein is covalently conjugated to target proteins upon activation of the interferon response. Interaction between vIRF1 and HERC5 was confirmed by immunoprecipitation, and the region between amino acids 224 and 349 of vIRF1 was required for interaction with HERC5. We further report that expression of vIRF1 in the context of TLR3 activation results in decreased ISG15 conjugation of proteins. Specifically, TLR3-induced ISG15 conjugation and protein levels of cellular IRF3, a known ISG15 target, were decreased in the presence of vIRF1 compared to the control. vIRF1 itself was also identified as a target of ISG15 conjugation. KSHV-infected cells exhibited increased ISG15 conjugation upon reactivation from latency in coordination with increased IFN. Furthermore, knockdown of ISG15 in latently infected cells resulted in a higher level of KSHV reactivation and an increase in infectious virus. These data suggest that the KSHV vIRF1 protein affects ISG15 conjugation and interferon responses and may contribute to effective KSHV replication. IMPORTANCEThe KSHV vIRF1 protein can inhibit interferon activation in response to viral infection. We identified a cellular protein named HERC5, which is the major ligase for ISG15, as a vIRF1 binding partner. vIRF1 association with HERC5 altered ISG15 modification of cellular proteins, and knockdown of ISG15 augmented reactivation of KSHV from latency. U pon detection of viral infection, cells activate the quintessential antiviral immune response, which results in production of type I interferon (IFN), including interferon alpha (IFN-␣) and IFN-␤. Type I IFN induces an antiviral transcriptional program, producing proteins that cooperate to inhibit the spread of infection. One of the most abundantly produced transcripts upon induction of the type I IFN response is interferon-stimulated gene 15 (ISG15) (1, 2). ISG15 is a ubiquitin-like 17-kDa protein that is covalently conjugated to target proteins via a process called ISGylation (3-5). Similar to ubiquitin, ISG15 requires three enzymatic steps for conjugation onto target proteins. They include the E1 enzyme Ube1L, the conjugating E2 enzyme UbcH8, and the major E3 ligase HERC5 (6). Multiple large-scale screens have identified hundreds of potential cellular targets of ISGylation; however, in most cases, the specific function of ISG15 conjugation to cellular proteins remains undetermined (7).Thus far, identified ISG15 target proteins function in a variety of cellular pathways, including glycolysis, cell motility, translat...

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Section: Discussionmentioning

confidence: 99%

Kaposi's Sarcoma-Associated Herpesvirus Viral Interferon Regulatory Factor 1 Interacts with a Member of the Interferon-Stimulated Gene 15 Pathway

Jacobs

Stopford

West

et al. 2015

J Virol

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“…11 Therefore, we evaluated the conjugation of p53 to SUMO2 in HEK-293 cells co-transfected with pcDNA, LANA2-WT, a LANA2 mutant in the SIM domain (LANA2DSIM), a SUMOylation mutant (LANA2DSUMO), or the double SIM and SUMOylation mutant of LANA2 (LANA2TOTAL), together with an empty vector or Ubc9 and His6-SUMO2. Expression of LANA2-WT induced a reduction in the levels of p53-SUMO2 that was less evident after expression of LANA2DSIM or LANA2DSUMO, and almost undetectable after expression of LANA2TOTAL ( Fig.…”

Section: Resultsmentioning

confidence: 99%

KSHV latent protein LANA2 inhibits sumo2 modification of p53

et al. 2015

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Tumor suppressor p53 plays a crucial antiviral role and targeting of p53 by viral proteins is a common mechanism involved in virus oncogenesis. The activity of p53 is tightly regulated at the post-translational levels through a myriad of modifications. Among them, modification of p53 by SUMO has been associated with the onset of cellular senescence. Kaposi´s sarcoma-associated herpesvirus (KSHV) expresses several proteins targeting p53, including the latent protein LANA2 that regulates polyubiquitylation and phosphorylation of p53. Here we show that LANA2 also inhibits the modification of p53 by SUMO2. Furthermore, we show that the reduction of p53-SUMO2 conjugation by LANA2, as well as the p53-LANA2 interaction, both require the SUMOylation of the viral protein and its interaction with SUMO or SUMOylated proteins in a non-covalent manner. Finally, we show that the control of p53-SUMO2 conjugation by LANA2 correlates with its ability to inhibit SUMO2-and type I interferon-induced senescence. These results highlight the importance of p53 SUMOylation in the control of virus infection and suggest that viral oncoproteins could contribute to viral infection and cell transformation by abrogating p53 SUMOylation.

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“…Viruses can counteract or enhance the host's sumoylation to evade the immune response and antagonize apoptosis, while some viruses exploit the host's sumoylation machinery to sumoylate the viral components which are necessary to perform viral functions (48). It has been reported that KSHV modulates the SUMO pathway through RTA/ORF50, K-bZIP/K8, and vPK/ORF36 during lytic replication as well as LANA/ORF73 and viral IRF3 (IRF3)/K10.5/LANA-2 during latency (27,35,(49)(50)(51)(52)(53)(54)(55)(56). Our results showed that LANA bound directly to the SENP6 promoter and repressed its activity, inhibiting SENP6 expression at both the RNA and protein levels.…”

Section: Discussionmentioning

confidence: 99%

The Latency-Associated Nuclear Antigen of Kaposi's Sarcoma-Associated Herpesvirus Inhibits Expression of SUMO/Sentrin-Specific Peptidase 6 To Facilitate Establishment of Latency

Lin

Sun

Zhang

et al. 2017

J Virol

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Kaposi's sarcoma-associated herpesvirus (KSHV), which belongs to the Gammaherpesviridae, typically displays two different phases in its life cycle, the latent phase and the lytic phase. Latency-associated nuclear antigen (LANA), the primary viral product during latency, has been reported to bind to a series of cellular gene promoters to modulate gene transcription. To systemically elucidate the cellular genes regulated by LANA, we identified genome-wide LANA binding sites by chromatin immunoprecipitation coupled with sequencing (ChIP-seq). We stratified ChIP-seq data and found that LANA might be involved in the macromolecule catabolic process. Specifically, we found and verified that LANA could directly bind to the promoter of the SUMO/sentrin-specific peptidase 6 (SENP6) gene in vivo and in vitro. LANA could repress SENP6 promoter activity in a dose-dependent manner in a reporter gene assay. LANA expression was sufficient to inhibit endogenous SENP6 expression at both the RNA and protein levels. Moreover, SENP6 overexpression in KSHV-infected cells reduced LANA at the protein level. Mechanistically, we found that SENP6 could interact with LANA and reduce the formation of sumoylated LANA, which relies on the desumoylation ability of SENP6. During de novo infection, SENP6 overexpression would decrease the abundance of LANA and enhance viral gene expression, which would hamper the establishment of latency. Taken together, these data suggest that KSHV-encoded LANA could inhibit SENP6 expression to regulate the abundance of itself, which may play an important role in controlling the establishment of latency.IMPORTANCE LANA, as a key latent protein produced by KSHV, is responsible for episome persistence and regulates viral reactivation. In the present study, our results demonstrated that LANA could bind to the promoter region of the SENP6 gene and inhibit SENP6 expression while the regulated SENP6 could in turn modulate the abundance of LANA through desumoylation. This delicate regulation may provide important insights to explain the abundance of LANA during KSHV latency.

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Covalent modification by SUMO is required for efficient disruption of PML oncogenic domains by Kaposi's sarcoma-associated herpesvirus latent protein LANA2

Cited by 32 publications

References 38 publications

Kaposi's Sarcoma-Associated Herpesvirus Viral Interferon Regulatory Factor 1 Interacts with a Member of the Interferon-Stimulated Gene 15 Pathway

Kaposi's Sarcoma-Associated Herpesvirus Viral Interferon Regulatory Factor 1 Interacts with a Member of the Interferon-Stimulated Gene 15 Pathway

KSHV latent protein LANA2 inhibits sumo2 modification of p53

The Latency-Associated Nuclear Antigen of Kaposi's Sarcoma-Associated Herpesvirus Inhibits Expression of SUMO/Sentrin-Specific Peptidase 6 To Facilitate Establishment of Latency

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