The Epstein-Barr Virus (EBV) Deubiquitinating Enzyme BPLF1 Reduces EBV Ribonucleotide Reductase Activity

Whitehurst, Christopher B.; Ning, Shunbin; Bentz, Gretchen L.; Dufour, Florent; Gershburg, Edward; Shackelford, Julia; Langelier, Yves; Pagano, Joseph S.

doi:10.1128/jvi.02195-08

Cited by 64 publications

(78 citation statements)

References 47 publications

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“…Polymorphisms in the genes encoding tegument and early lytic proteins and glycoproteins have been identified here. BPLF1 is one of the few tegument proteins whose function has been relatively well investigated (41). We identified two nonsynonymous mutations in the N-terminal region of the protein that exhibits deubiquitinating activity.…”

Section: Discussionmentioning

confidence: 98%

Genomic Diversity of Epstein-Barr Virus Genomes Isolated from Primary Nasopharyngeal Carcinoma Biopsy Samples

Kwok

Palser

et al. 2014

J Virol

101

111

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Undifferentiated nasopharyngeal carcinoma (NPC) has a 100% association with Epstein-Barr virus (EBV). However, only three EBV genomes isolated from NPC patients have been sequenced to date, and the role of EBV genomic variations in the pathogenesis of NPC is unclear. We sought to obtain the sequences of EBV genomes in multiple NPC biopsy specimens in the same geographic location in order to reveal their sequence diversity. Three published EBV (B95-8, C666-1, and HKNPC1) genomes were first resequenced using the sequencing workflow of target enrichment of EBV DNA by hybridization, followed by next-generation sequencing, de novo assembly, and joining of contigs by Sanger sequencing. The sequences of eight NPC biopsy specimenderived EBV (NPC-EBV) genomes, designated HKNPC2 to HKNPC9, were then determined. They harbored 1,736 variations in total, including 1,601 substitutions, 64 insertions, and 71 deletions, compared to the reference EBV. Furthermore, genes encoding latent, early lytic, and tegument proteins and glycoproteins were found to contain nonsynonymous mutations of potential biological significance. Phylogenetic analysis showed that the HKNPC6 and -7 genomes, which were isolated from tumor biopsy specimens of advanced metastatic NPC cases, were distinct from the other six NPC-EBV genomes, suggesting the presence of at least two parental lineages of EBV among the NPC-EBV genomes. In conclusion, much greater sequence diversity among EBV isolates derived from NPC biopsy specimens is demonstrated on a whole-genome level through a complete sequencing workflow. Large-scale sequencing and comparison of EBV genomes isolated from NPC and normal subjects should be performed to assess whether EBV genomic variations contribute to NPC pathogenesis. IMPORTANCEThis study established a sequencing workflow from EBV DNA capture and sequencing to de novo assembly and contig joining. We reported eight newly sequenced EBV genomes isolated from primary NPC biopsy specimens and revealed the sequence diversity on a whole-genome level among these EBV isolates. At least two lineages of EBV strains are observed, and recombination among these lineages is inferred. Our study has demonstrated the value of, and provided a platform for, genome sequencing of EBV.T he incidence rate of undifferentiated nasopharyngeal carcinoma (NPC) is exceptionally high in the southern part of China, and this type of carcinoma is 100% associated with Epstein-Barr virus (EBV) (1). To investigate the role of EBV genomic variation in the pathogenesis of NPC, EBV strains had been characterized in NPC by genotyping polymorphic markers in the EBER1 and -2, LMP1, BHRF1, BZLF1, and EBNA1 gene loci in tumor samples obtained from China, southern Asia, and northern Africa (2-7). Association of LMP1 deletion variant Asp335 with NPC in Hong Kong was reported (8). Specific EBNA1 (V-val) and LMP1 subtypes (China 1) also showed preferential occurrence in NPC biopsy specimens (9, 10). However, genetic variations in the small subsets of genes investigated were not ...

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Section: Discussionmentioning

confidence: 98%

Genomic Diversity of Epstein-Barr Virus Genomes Isolated from Primary Nasopharyngeal Carcinoma Biopsy Samples

Kwok

Palser

et al. 2014

J Virol

101

111

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“…The calcium phosphate technique [7] was used to transfect 10 lg per well and 50 lg per dish of the plasmids pAdCMV5 (empty), pAdCMV5-R1 (HSV-2 R1); pLBPF1-GST (GST), pLBPF1-GST-R1 (GST-HSV-1 R1) (kindly provided by A. Pearson); pcDNA3 (empty), pcDNA3-HA-EBV R1 (HA-EBV R1) [29]; pEGFP C1 (GFP) and plasmids encoding for caspase-8 GFP (casp-8 GFP), caspase-8 C360S GFP (casp-8 C360S GFP), caspase-8 DED-AB (1-209) GFP (casp-8 DED-AB GFP), caspase-8 CD (210-479) GFP (casp-8 CD GFP) and FADD YFP [30]. Immunofluorescence was performed on cells fixed with paraformaldehyde as previously described [31].…”

Section: Transfection and Immunofluorescencementioning

confidence: 99%

The ribonucleotide reductase R1 subunits of herpes simplex virus types 1 and 2 protect cells against TNFα- and FasL-induced apoptosis by interacting with caspase-8

et al. 2010

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We previously reported that HSV-2 R1, the R1 subunit (ICP10; UL39) of herpes simplex virus type-2 ribonucleotide reductase, protects cells against apoptosis induced by the death receptor (DR) ligands tumor necrosis factor-alpha-(TNFa) and Fas ligand (FasL) by interrupting DR-mediated signaling at, or upstream of, caspase-8 activation. Further investigation of the molecular mechanism underlying HSV-2 R1 protection showed that extracellularregulated kinase 1/2 (ERK1/2), phosphatidylinositol 3-kinase (PI3-K)/Akt, NF-jB and JNK survival pathways do not play a major role in this antiapoptotic function. Interaction studies revealed that HSV-2 R1 interacted constitutively with caspase-8. The HSV-2 R1 deletion mutant R1(1-834)-GFP and Epstein-Barr virus (EBV) R1, which did not protect against apoptosis induced by DR ligands, did not interact with caspase-8, indicating that interaction is required for protection. HSV-2 R1 impaired caspase-8 activation induced by caspase-8 over-expression, suggesting that interaction between the two proteins prevents caspase-8 dimerization/activation. HSV-2 R1 bound to caspase-8 directly through its prodomain but did not interact with either its caspase domain or Fas-associated death domain protein (FADD). Interaction between HSV-2 R1 and caspase-8 disrupted FADD-caspase-8 binding. We further demonstrated that individually expressed HSV-1 R1 (ICP6) shares, with HSV-2 R1, the ability to bind caspase-8 and to protect cells against DR-induced apoptosis. Finally, as the long-lived Fas protein remained stable during the early period of infection, experiments with the HSV-1 UL39 deletion mutant ICP6D showed that HSV-1 R1 could be essential for the protection of HSV-1-infected cells against FasL.Keywords FasL Á ICP6 Á ICP10 Á Viral inhibitor of apoptosis Á Caspase-8Electronic supplementary material The online version of this article

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“…Recently, a number of studies have shown that herpesviridae have large tegument proteins with a DUB domain embedded at their N terminus, such as herpes simplex virus 1 (HSV-1) UL36 USP , mouse cytomegalovirus (MCMV) M48, human cytomegalovirus (HCMV) UL48, EBV BPLF1, and KSHV and MHV68 ORF64 (11,14,25,26,31). These viral tegument DUBs can cleave either Lys48-or Lys63-linked polyubiquitin chains in in vitro deubiquitination assays (11,16,26,33). Since ubiquitination plays a critical role in RIG-I signaling (10), we tested whether KSHV ORF64 inhibits RIG-I function.…”

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confidence: 99%

Inhibition of RIG-I-Mediated Signaling by Kaposi's Sarcoma-Associated Herpesvirus-Encoded Deubiquitinase ORF64

Inn

Lee

Rathbun

et al. 2011

J Virol

144

150

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The Epstein-Barr Virus (EBV) Deubiquitinating Enzyme BPLF1 Reduces EBV Ribonucleotide Reductase Activity

Cited by 64 publications

References 47 publications

Genomic Diversity of Epstein-Barr Virus Genomes Isolated from Primary Nasopharyngeal Carcinoma Biopsy Samples

Genomic Diversity of Epstein-Barr Virus Genomes Isolated from Primary Nasopharyngeal Carcinoma Biopsy Samples

The ribonucleotide reductase R1 subunits of herpes simplex virus types 1 and 2 protect cells against TNFα- and FasL-induced apoptosis by interacting with caspase-8

Inhibition of RIG-I-Mediated Signaling by Kaposi's Sarcoma-Associated Herpesvirus-Encoded Deubiquitinase ORF64

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