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The Epstein-Barr virus (EBV) nuclear antigen (EBNA)-1 is the only viral protein expressed in all EBV-carrying malignancies, but its contribution to oncogenesis has remained enigmatic. We show that EBNA-1 induces chromosomal aberrations, DNA double-strand breaks, and engagement of the DNA damage response (DDR). These signs of genomic instability are associated with the production of reactive oxygen species (ROS) and are reversed by antioxidants. The catalytic subunit of the leukocyte NADPH oxidase, NOX2/gp91 phox , is transcriptionally activated in EBNA-1-expressing cells, whereas inactivation of the enzyme by chemical inhibitors or RNAi halts ROS production and DDR. These findings highlight a novel function of EBNA-1 and a possible mechanism by which expression of this viral protein could contribute to malignant transformation and tumor progression.is a human gamma-herpesvirus that establishes latent infections in B lymphocytes, where only a subset of viral genes is expressed and virus replication is suppressed (1). The proteins encoded by the latency genes, including 6 EBV-encoded nuclear antigens (EBNA-1, -2, -3A, -3B, -3C, and -5) and 3 latent membrane proteins (LMP1, -2A, and -2B), induce growth transformation by capturing multiple signaling pathways that control B cell proliferation and apoptosis. It is generally assumed that the continuous expression of viral genes underlies the association of EBV with a variety of human malignancies, including Burkitt's lymphoma (BL), Hodgkin's disease (HD), nasopharyngeal carcinoma (NPC), and posttransplant lymphoproliferative disease (PTLD) (2). Some EBVpositive tumors do not express all of the latency proteins, leading to restricted forms of latency in which EBNA-1 is detected either alone (latency I, found in BL) or together with the LMPs (latency II, found in HD and NPC). Thus, EBNA-1 is the only viral protein regularly expressed in all EBV-carrying malignancies.EBNA-1 binds to the viral origin of replication (oriP) and is required for the correct partitioning of the viral episomes in proliferating cells (3). It may confer a growth advantage to BL cells (4) and protect them from apoptosis (5) but does not act as an autonomous oncogene (6) and seems to be dispensable for B cell immortalization in vitro (7). Hence, the mechanism by which EBNA-1 may contribute to malignant transformation is not understood.Genomic instability is common in malignant cells and was observed in EBV-carrying tumors (8-10). EBNA-3C (11) and LMP-1 (12) may promote this phenotype through inhibition of DNA repair or inactivation of cell cycle checkpoints, which allow the propagation of DNA damage. However, these viral proteins are not expressed in EBV-carrying BLs, and only half of HDs and NPCs express detectable levels of LMP1, suggesting a limited role in EBV oncogenesis. A possible involvement of EBNA-1 in the induction of genomic instability is suggested by a significant increase of transient chromosomal aberrations, such as dicentric chromosomes, chromosome fragments, and gaps, in EBVpositiv...

SUMMARY The human gut is colonized by a large number of microorganisms (~1013 bacteria) that support various physiologic functions. A perturbation in healthy gut microbiome might leads to the development of inflammatory diseases including multiple sclerosis (MS). Therefore, gut commensals can provide promising therapeutic options for treating autoimmune diseases such as MS. We report identification of human gut–derived commensal bacteria, Prevotella histicola, which can suppress an autoimmune disease in HLA class-II transgenic model of experimental autoimmune encephalomyelitis (EAE); an animal model of MS. P. histicola suppresses disease through modulation of systemic immune responses. P. histicola challenge led to a decrease in pro-inflammatory Th1 and Th17 cells, and increase in the frequencies of CD4+FoxP3+ regulatory T cells, tolerogenic dendritic cells, and suppressive macrophage. Our study provides evidence that administration of gut commensals may regulate a systemic immune response and may, therefore, have a possible role in the treatment strategies for MS.

Ubiquitination regulates membrane events such as endocytosis, membrane trafficking and endoplasmic-reticulum-associated degradation (ERAD). Although the involvement of membraneassociated ubiquitin-conjugating enzymes and ligases in these processes is well documented, their regulation by ubiquitin deconjugases is less well understood. By screening a database of human deubiquitinating enzymes (DUBs), we have identified a putative transmembrane domain in ubiquitin-specific protease (USP)19. We show that USP19 is a tail-anchored ubiquitin-specific protease localized to the ER and is a target of the unfolded protein response. USP19 rescues the ERAD substrates cystic fibrosis transmembrane conductance regulator (CFTR)DF508 and T-cell receptor-a (TCRa) from proteasomal degradation. A catalytically inactive USP19 was still able to partly rescue TCRa but not CFTRDF508, suggesting that USP19 might also exert a non-catalytic function on specific ERAD substrates. Thus, USP19 is the first example of a membrane-anchored DUB involved in the turnover of ERAD substrates.

Mutations in the gene encoding transcription factor TFAP2A result in pigmentation anomalies in model organisms and premature hair graying in humans. However, the pleiotropic functions of TFAP2A and its redundantly-acting paralogs have made the precise contribution of TFAP2-type activity to melanocyte differentiation unclear. Defining this contribution may help to explain why TFAP2A expression is reduced in advanced-stage melanoma compared to benign nevi. To identify genes with TFAP2A-dependent expression in melanocytes, we profile zebrafish tissue and mouse melanocytes deficient in Tfap2a, and find that expression of a small subset of genes underlying pigmentation phenotypes is TFAP2A-dependent, including Dct, Mc1r, Mlph, and Pmel. We then conduct TFAP2A ChIP-seq in mouse and human melanocytes and find that a much larger subset of pigmentation genes is associated with active regulatory elements bound by TFAP2A. These elements are also frequently bound by MITF, which is considered the “master regulator” of melanocyte development. For example, the promoter of TRPM1 is bound by both TFAP2A and MITF, and we show that the activity of a minimal TRPM1 promoter is lost upon deletion of the TFAP2A binding sites. However, the expression of Trpm1 is not TFAP2A-dependent, implying that additional TFAP2 paralogs function redundantly to drive melanocyte differentiation, which is consistent with previous results from zebrafish. Paralogs Tfap2a and Tfap2b are both expressed in mouse melanocytes, and we show that mouse embryos with Wnt1-Cre-mediated deletion of Tfap2a and Tfap2b in the neural crest almost completely lack melanocytes but retain neural crest-derived sensory ganglia. These results suggest that TFAP2 paralogs, like MITF, are also necessary for induction of the melanocyte lineage. Finally, we observe a genetic interaction between tfap2a and mitfa in zebrafish, but find that artificially elevating expression of tfap2a does not increase levels of melanin in mitfa hypomorphic or loss-of-function mutants. Collectively, these results show that TFAP2 paralogs, operating alongside lineage-specific transcription factors such as MITF, directly regulate effectors of terminal differentiation in melanocytes. In addition, they suggest that TFAP2A activity, like MITF activity, has the potential to modulate the phenotype of melanoma cells.