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Article ; Online: Factors influencing stoichiometry and stability of polyoxometalate - peptide complexes.

Greijer, Björn H / Nestor, Gustav / Eriksson, Jan E / Seisenbaeva, Gulaim A / Kessler, Vadim G

Dalton transactions (Cambridge, England : 2003)

2022 Volume 51, Issue 24, Page(s) 9511–9521

Abstract: In the pursuit of understanding the factors guiding interactions between polyoxometalates (POMs) and biomolecules, several complexes between Keggin phosphomolybdate and diglycine have been produced at different acidity and salinity conditions, leading to ...

Abstract	In the pursuit of understanding the factors guiding interactions between polyoxometalates (POMs) and biomolecules, several complexes between Keggin phosphomolybdate and diglycine have been produced at different acidity and salinity conditions, leading to difference in stoichiometry and in crystal structure. Principal factors determining how the POM and dipeptide interact appear to be pH, ionic strength of the medium, and the molar ratio of POM to peptide. An important effect turned out to be even the structure-directing role of the sodium cations coordinating carbonyl functions of the peptide bond. Given the interest in applying POMs in biological systems, these factors are highly relevant to consider. In the view of recent interest in using POMs as nano catalysts in peptide hydrolysis also the potential Keggin POM transformation in phosphate buffered saline medium was investigated leading to insight that nanoparticles of zirconium phosphate (ZrP) can be actual catalysts for breakdown of the peptide bond.
MeSH term(s)	Anions ; Hydrolysis ; Peptides/chemistry ; Polyelectrolytes ; Tungsten Compounds/chemistry
Chemical Substances	Anions ; Peptides ; Polyelectrolytes ; Tungsten Compounds ; polyoxometalate
Language	English
Publishing date	2022-06-21
Publishing country	England
Document type	Journal Article
ZDB-ID	1472887-4
ISSN	1477-9234 ; 1364-5447 ; 0300-9246 ; 1477-9226
ISSN (online)	1477-9234 ; 1364-5447
ISSN	0300-9246 ; 1477-9226
DOI	10.1039/d2dt00717g
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Article: Epstein-Barr Virus Gene BARF1 Expression is Regulated by the Epithelial Differentiation Factor ΔNp63α in Undifferentiated Nasopharyngeal Carcinoma.

Hoebe, Eveline / Wille, Coral / Hagemeier, Stacy / Kenney, Shannon / Greijer, Astrid / Middeldorp, Jaap

Cancers

2018 Volume 10, Issue 3

Abstract: Epstein-Barr Virus (EBV) BamHI-A rightward frame 1 (BARF1) protein is considered a viral oncogene in epithelial cells and has immune-modulating properties. During viral lytic replication BARF1 is expressed as an early gene, regulated by the immediate ... ...

Abstract	Epstein-Barr Virus (EBV) BamHI-A rightward frame 1 (BARF1) protein is considered a viral oncogene in epithelial cells and has immune-modulating properties. During viral lytic replication BARF1 is expressed as an early gene, regulated by the immediate early EBV protein R. However, in viral latency BARF1 is exclusively expressed in epithelial tumors such as nasopharyngeal (NPC) and gastric carcinoma (GC) but not in lymphomas, indicating that activation of the BARF1 promoter is cell type specific. Undifferentiated NPC is characterized by high expression of ΔNp63 isoforms of the epithelial differentiation marker p63, a member of the p53 family of transcription factors. Transcription factor binding site analysis indicated potential p53 family binding sites within the BARF1 promoter region. This study investigated ability of various p53 family members to transactivate the BARF1 promoter. Using BARF1 promoter luciferase reporter constructs we demonstrate that only p63 isoform ΔNp63α is capable of transactivating the BARF1 promoter, but not the TAp63 isoforms, p53 or p73. Direct promoter binding of ΔNp63α was confirmed by Chromatin Immune Precipitation (ChIP) analysis. Deletion mutants of the BARF1 promoter revealed multiple ΔNp63 response elements to be responsible for BARF1 promoter transactivation. However, ΔNp63α alone was not sufficient to induce BARF1 in tumor cells harboring full EBV genomes, indicating that additional cofactors might be required for full BARF1 regulation. In conclusion, in EBV positive NPC and GC, BARF1 expression might be induced by the epithelial differentiation marker ΔNp63α, explaining BARF1 expression in the absence of lytic reactivation.
Language	English
Publishing date	2018-03-17
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527080-1
ISSN	2072-6694
ISSN	2072-6694
DOI	10.3390/cancers10030076
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Article: Quantitative multi-target RNA profiling in Epstein-Barr virus infected tumor cells

Greijer, A.E / H. Juwana / J.M. Middeldorp / O. Ramayanti / S.A.W.M. Verkuijlen / Z. NovaliÄ

Journal of virological methods. 2017 Mar., v. 241

2017

Abstract: Epstein-Barr virus (EBV) is etiologically linked to multiple acute, chronic and malignant diseases. Detection of EBV-RNA transcripts in tissues or biofluids besides EBV-DNA can help in diagnosing EBV related syndromes. Sensitive EBV transcription ... ...

Abstract	Epstein-Barr virus (EBV) is etiologically linked to multiple acute, chronic and malignant diseases. Detection of EBV-RNA transcripts in tissues or biofluids besides EBV-DNA can help in diagnosing EBV related syndromes. Sensitive EBV transcription profiling yields new insights on its pathogenic role and may be useful for monitoring virus targeted therapy. Here we describe a multi-gene quantitative RT-PCR profiling method that simultaneously detects a broad spectrum (n=16) of crucial latent and lytic EBV transcripts. These transcripts include (but are not restricted to), EBNA1, EBNA2, LMP1, LMP2, BARTs, EBER1, BARF1 and ZEBRA, Rta, BGLF4 (PK), BXLF1 (TK) and BFRF3 (VCAp18) all of which have been implicated in EBV-driven oncogenesis and viral replication. With this method we determine the amount of RNA copies per infected (tumor) cell in bulk populations of various origin. While we confirm the expected RNA profiles within classic EBV latency programs, this sensitive quantitative approach revealed the presence of rare cells undergoing lytic replication. Inducing lytic replication in EBV tumor cells supports apoptosis and is considered as therapeutic approach to treat EBV-driven malignancies. This sensitive multi-primed quantitative RT-PCR approach can provide broader understanding of transcriptional activity in latent and lytic EBV infection and is suitable for monitoring virus-specific therapy responses in patients with EBV associated cancers.
Keywords	apoptosis ; carcinogenesis ; Human herpesvirus 4 ; monitoring ; neoplasm cells ; neoplasms ; patients ; reverse transcriptase polymerase chain reaction ; RNA ; therapeutics ; tissues ; transcription (genetics) ; virus replication ; viruses
Language	English
Dates of publication	2017-03
Size	p. 24-33.
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	8013-5
ISSN	1879-0984 ; 0166-0934
ISSN (online)	1879-0984
ISSN	0166-0934
DOI	10.1016/j.jviromet.2016.12.007
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Article ; Online: Quantitative multi-target RNA profiling in Epstein-Barr virus infected tumor cells.

Greijer, A E / Ramayanti, O / Verkuijlen, S A W M / Novalić, Z / Juwana, H / Middeldorp, J M

Journal of virological methods

2016 Volume 241, Page(s) 24–33

Abstract	Epstein-Barr virus (EBV) is etiologically linked to multiple acute, chronic and malignant diseases. Detection of EBV-RNA transcripts in tissues or biofluids besides EBV-DNA can help in diagnosing EBV related syndromes. Sensitive EBV transcription profiling yields new insights on its pathogenic role and may be useful for monitoring virus targeted therapy. Here we describe a multi-gene quantitative RT-PCR profiling method that simultaneously detects a broad spectrum (n=16) of crucial latent and lytic EBV transcripts. These transcripts include (but are not restricted to), EBNA1, EBNA2, LMP1, LMP2, BARTs, EBER1, BARF1 and ZEBRA, Rta, BGLF4 (PK), BXLF1 (TK) and BFRF3 (VCAp18) all of which have been implicated in EBV-driven oncogenesis and viral replication. With this method we determine the amount of RNA copies per infected (tumor) cell in bulk populations of various origin. While we confirm the expected RNA profiles within classic EBV latency programs, this sensitive quantitative approach revealed the presence of rare cells undergoing lytic replication. Inducing lytic replication in EBV tumor cells supports apoptosis and is considered as therapeutic approach to treat EBV-driven malignancies. This sensitive multi-primed quantitative RT-PCR approach can provide broader understanding of transcriptional activity in latent and lytic EBV infection and is suitable for monitoring virus-specific therapy responses in patients with EBV associated cancers.
MeSH term(s)	Biomarkers ; Epstein-Barr Virus Nuclear Antigens/genetics ; Gene Expression Profiling/methods ; Herpesvirus 4, Human/genetics ; Herpesvirus 4, Human/physiology ; Humans ; RNA, Messenger/analysis ; RNA, Messenger/genetics ; RNA, Viral/genetics ; Tumor Cells, Cultured ; Viral Matrix Proteins/genetics ; Viral Proteins/genetics ; Virus Latency/genetics ; Virus Replication
Chemical Substances	Biomarkers ; Epstein-Barr Virus Nuclear Antigens ; Epstein-Barr virus encoded RNA 1 ; RNA, Messenger ; RNA, Viral ; Viral Matrix Proteins ; Viral Proteins ; EBV-encoded nuclear antigen 1 (O5GA75RST7)
Language	English
Publishing date	2016-12-16
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	8013-5
ISSN	1879-0984 ; 0166-0934
ISSN (online)	1879-0984
ISSN	0166-0934
DOI	10.1016/j.jviromet.2016.12.007
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Article ; Online: Epstein-Barr virus mRNA profiles and viral DNA methylation status in nasopharyngeal brushings from nasopharyngeal carcinoma patients reflect tumor origin.

Ramayanti, Octavia / Juwana, Hedy / Verkuijlen, Sandra A M W / Adham, Marlinda / Pegtel, Michiel D / Greijer, Astrid E / Middeldorp, Jaap M

International journal of cancer

2016 Volume 140, Issue 1, Page(s) 149–162

Abstract: Undifferentiated nasopharyngeal carcinoma (NPC) is 100% associated with Epstein-Barr virus (EBV) as oncogenic driver. NPC is often diagnosed late due to initial vague complaints and obscured location. Prior studies suggest that measurement of EBV DNA ... ...

Abstract	Undifferentiated nasopharyngeal carcinoma (NPC) is 100% associated with Epstein-Barr virus (EBV) as oncogenic driver. NPC is often diagnosed late due to initial vague complaints and obscured location. Prior studies suggest that measurement of EBV DNA load and RNA transcripts in nasopharyngeal (NP) brushings is useful for minimally invasive NPC diagnosis. However, whether these EBV markers relate to local virus replication or reflect tumor origin remains to be demonstrated. To resolve this, we analysed EBV-DNA characteristics and quantified latent and lytic viral RNA transcripts in NP brushings and matching frozen NP-biopsy specimens from patients suspected of having NPC. We observed non-fragmented and Cp-promotor methylated EBV-DNA in both NP brushings and biopsies suggestive of tumor origin. Using quantitative RT-PCR we determined expression levels of 7 critical latent (EBER1, Qp-EBNA1, EBNA2, BART, LMP1, LMP2, BARF1) and 5 lytic (Zta, Rta, TK, PK and VCA-p18) RNA transcripts. Although latent and early lytic RNA transcripts were frequently detected in conjunction with high EBV viral load, in both brushings and biopsies the latent transcripts prevailed and reflected a typical NPC-associated latency-II transcription profile without EBNA2. Late lytic RNA transcripts were rare and detected at low levels mainly in NP brushings, suggestive of abortive viral reactivation rather than complete virus replication. EBV-IgA serology (EBNA1, VCA, Zta) did not correlate to the level of viral reactivation in situ. Overall, viral RNA profiling, DNA fragmentation and methylation analysis in NP brushings and parallel biopsies indicate that NP brush sampling provides a true and robust indicator of NPC tumor presence.
MeSH term(s)	Adult ; Biopsy ; Carcinoma ; DNA Methylation ; DNA, Viral/genetics ; Early Detection of Cancer ; Epstein-Barr Virus Infections/genetics ; Female ; Gene Expression Profiling/methods ; Gene Expression Regulation, Viral ; Herpesvirus 4, Human/genetics ; Herpesvirus 4, Human/physiology ; Humans ; Male ; Middle Aged ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms/diagnosis ; Nasopharyngeal Neoplasms/pathology ; Nasopharyngeal Neoplasms/virology ; Nasopharynx/virology ; Promoter Regions, Genetic ; Prospective Studies ; RNA, Messenger/genetics ; RNA, Viral/genetics ; Viral Load
Chemical Substances	DNA, Viral ; RNA, Messenger ; RNA, Viral
Language	English
Publishing date	2016-09-23
Publishing country	United States
Document type	Journal Article
ZDB-ID	218257-9
ISSN	1097-0215 ; 0020-7136
ISSN (online)	1097-0215
ISSN	0020-7136
DOI	10.1002/ijc.30418
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Zs.A 478: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Epstein-Barr DNA in advanced pediatric nasopharyngeal cancer

Marlinda Adham / Namira Kesuma Jelita / Djajadiman Gatot / Soehartati Argadikoesoema Gondhowiardjo / Lisnawati Rachmadi / Astrid E Greijer / I Bing Tan / Jaap M Middeldorp

Paediatrica Indonesiana, Vol 61, Iss 5, Pp 261-

2021 Volume 70

Abstract: Background Studies suggest that the most common type of nasopharyngeal carcinoma (NPC) is WHO-3, which is strongly associated with Epstein-Barr virus (EBV). Objective To assess NPC patient characteristics in a national general referral hospital in ... ...

Abstract	Background Studies suggest that the most common type of nasopharyngeal carcinoma (NPC) is WHO-3, which is strongly associated with Epstein-Barr virus (EBV). Objective To assess NPC patient characteristics in a national general referral hospital in Indonesia, with regards to EBV DNA load and treatment response. Methods Twenty-three pediatric patients diagnosed with NPC were included in the study. Data collected were history, physical examination, tissue biopsy, CT scan, staging and EBV DNA load from nasopharyngeal (NP) brushing as well as blood specimens. The NP brushing, blood specimens and CT scan evaluations were done two months post-treatment. Results Pediatric patients with symptoms such as blood tinged secretion, lymph node enlargement, and nasal congestion were more likely to have higher EBV DNA loads in their NP brushings (P<0.05) (including T3 and higher). Despite significant reduction of EBV DNA load in NP brushing post-treatment, it was not associated with treatment response, as evaluated by CT scan. Conclusion Higher DNA load from NP brushings is associated with a higher tumor stage. Larger sample size and follow-up data are needed to assess the usefulness of EBV DNA load assessment in pediatric patients.
Keywords	ebv ; pediatrics ; nasopharyngeal carcinoma ; asia ; indonesia ; Medicine ; R ; RJ1-570
Subject code	610
Language	English
Publishing date	2021-09-01T00:00:00Z
Publisher	Indonesian Pediatric Society Publishing House
Document type	Article ; Online
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Article ; Online: BamHI-A rightward frame 1, an Epstein-Barr virus-encoded oncogene and immune modulator.

Hoebe, Eveline K / Le Large, Tessa Y S / Greijer, Astrid E / Middeldorp, Jaap M

Reviews in medical virology

2013 Volume 23, Issue 6, Page(s) 367–383

Abstract: Epstein-Barr virus (EBV) causes several benign and malignant disorders of lymphoid and epithelial origin. EBV-related tumors display distinct patterns of viral latent gene expression, of which the BamHI-A rightward frame 1 (BARF1) is selectively ... ...

Abstract	Epstein-Barr virus (EBV) causes several benign and malignant disorders of lymphoid and epithelial origin. EBV-related tumors display distinct patterns of viral latent gene expression, of which the BamHI-A rightward frame 1 (BARF1) is selectively expressed in carcinomas, regulated by cellular differentiation factors including ΔNp63α. BARF1 functions as a viral oncogene, immortalizing and transforming epithelial cells of different origin by acting as a mitogenic growth factor, inducing cyclin-D expression, and up-regulating antiapoptotic Bcl-2, stimulating host cell growth and survival. In addition, secreted hexameric BARF1 has immune evasive properties, functionally corrupting macrophage colony stimulating factor, as supported by recent functional and structural data. Therefore, BARF1, an intracellular and secreted protein, not only has multiple pathogenic functions but also can function as a target for immune responses. Deciphering the role of BARF1 in EBV biology will contribute to novel diagnostic and treatment options for EBV-driven carcinomas. Herein, we discuss recent insights on the regulation of BARF1 expression and aspects of structure-function relating to its oncogenic and immune suppressive properties.
MeSH term(s)	Cell Transformation, Viral ; Herpesvirus 4, Human/immunology ; Herpesvirus 4, Human/physiology ; Host-Pathogen Interactions ; Humans ; Immune Evasion ; Oncogene Proteins, Viral/immunology ; Oncogene Proteins, Viral/metabolism ; Viral Proteins/immunology ; Viral Proteins/metabolism
Chemical Substances	BARF1 protein, Human herpesvirus 4 ; Oncogene Proteins, Viral ; Viral Proteins
Language	English
Publishing date	2013-08-31
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	1086043-5
ISSN	1099-1654 ; 1052-9276
ISSN (online)	1099-1654
ISSN	1052-9276
DOI	10.1002/rmv.1758
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Zs.A 3308: Show issues

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Article ; Online: Cytolytic virus activation therapy and treatment monitoring for Epstein-Barr virus associated nasopharyngeal carcinoma in a mouse tumor model.

Novalić, Zlata / Verkuijlen, Sandra A W M / Verlaan, Mariska / Eersels, Jos L H / de Greeuw, Inge / Molthoff, Carla F M / Middeldorp, Jaap M / Greijer, Astrid E

Journal of medical virology

2017 Volume 89, Issue 12, Page(s) 2207–2216

Abstract: Undifferentiated nasopharyngeal carcinoma (NPC) is 100% associated with Epstein-Barr virus (EBV). Expression of viral proteins in the tumor cells is highly restricted. EBV reactivation by CytoLytic Virus Activation (CLVA) therapy triggers de novo ... ...

Abstract	Undifferentiated nasopharyngeal carcinoma (NPC) is 100% associated with Epstein-Barr virus (EBV). Expression of viral proteins in the tumor cells is highly restricted. EBV reactivation by CytoLytic Virus Activation (CLVA) therapy triggers de novo expression of early viral kinases (PK and TK) and uses antiviral treatment to kill activated cells. The mechanism of tumor elimination by CLVA was analyzed in NPC mouse model using C666.1 cells. Valproic acid (VPA) was combined with gemcitabine (GCb) to stimulate EBV reactivation, followed by antiviral treatment with ganciclovir (GCV). A single cycle of CLVA treatment resulted in specific tumor cell killing as indicated by reduced tumor volume, loss of EBV-positive cells in situ, and paralleled by decreased EBV DNA levels in circulation, which was more pronounced than treatment with GCb alone. In vivo reactivation was confirmed by presence of lytic gene transcripts and proteins in tumors 6 days after GCb/VPA treatment. Virus reactivation was visualized by [
MeSH term(s)	Animals ; Antiviral Agents/blood ; Antiviral Agents/pharmacology ; Antiviral Agents/therapeutic use ; Carcinoma/drug therapy ; Carcinoma/virology ; DNA, Viral/genetics ; Deoxycytidine/analogs & derivatives ; Deoxycytidine/pharmacology ; Disease Models, Animal ; Epstein-Barr Virus Infections/complications ; Epstein-Barr Virus Infections/drug therapy ; Epstein-Barr Virus Infections/virology ; Ganciclovir/administration & dosage ; Ganciclovir/blood ; Ganciclovir/therapeutic use ; Herpesvirus 4, Human/drug effects ; Herpesvirus 4, Human/physiology ; Humans ; Mice ; Mice, Nude ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms/drug therapy ; Nasopharyngeal Neoplasms/virology ; Treatment Outcome ; Tumor Cells, Cultured ; Valproic Acid/pharmacology ; Valproic Acid/therapeutic use ; Viral Load/methods ; Virus Activation
Chemical Substances	Antiviral Agents ; DNA, Viral ; Deoxycytidine (0W860991D6) ; Valproic Acid (614OI1Z5WI) ; gemcitabine (B76N6SBZ8R) ; Ganciclovir (P9G3CKZ4P5)
Language	English
Publishing date	2017-08-29
Publishing country	United States
Document type	Journal Article
ZDB-ID	752392-0
ISSN	1096-9071 ; 0146-6615
ISSN (online)	1096-9071
ISSN	0146-6615
DOI	10.1002/jmv.24870
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Article: The role of hypoxia inducible factor 1 (HIF-1) in hypoxia induced apoptosis.

Greijer, A E / van der Wall, E

Journal of clinical pathology

2004 Volume 57, Issue 10, Page(s) 1009–1014

Abstract: Apoptosis can be induced in response to hypoxia. The severity of hypoxia determines whether cells become apoptotic or adapt to hypoxia and survive. A hypoxic environment devoid of nutrients prevents the cell undergoing energy dependent apoptosis and ... ...

Abstract	Apoptosis can be induced in response to hypoxia. The severity of hypoxia determines whether cells become apoptotic or adapt to hypoxia and survive. A hypoxic environment devoid of nutrients prevents the cell undergoing energy dependent apoptosis and cells become necrotic. Apoptosis regulatory proteins are delicately balanced. In solid tumours, hypoxia is a common phenomenon. Cells adapt to this environmental stress, so that after repeated periods of hypoxia, selection for resistance to hypoxia induced apoptosis occurs. These resistant tumours probably have a more aggressive phenotype and may have decreased responsiveness to treatment. The key regulator of this process, hypoxia inducible factor 1 (HIF-1), can initiate apoptosis by inducing high concentrations of proapoptotic proteins, such as BNIP3, and can cause stabilisation of p53. However, during hypoxia, antiapoptotic proteins, such as IAP-2, can be induced, whereas the proapoptotic protein Bax can be downregulated. During hypoxia, an intricate balance exists between factors that induce or counteract apoptosis, or even stimulate proliferation. Understanding the regulation of apoptosis during hypoxia and the mechanisms of resistance to apoptosis might lead to more specific treatments for solid tumours.
MeSH term(s)	Animals ; Apoptosis ; Cell Hypoxia/physiology ; DNA-Binding Proteins/physiology ; Gene Expression Regulation, Neoplastic ; Genes, p53 ; Humans ; Hypoxia-Inducible Factor 1 ; Hypoxia-Inducible Factor 1, alpha Subunit ; Inhibitor of Apoptosis Proteins ; Membrane Proteins/metabolism ; Necrosis ; Neoplasms/metabolism ; Neoplasms/pathology ; Nuclear Proteins/physiology ; Proteins/metabolism ; Proto-Oncogene Proteins/metabolism ; Proto-Oncogene Proteins c-bcl-2/metabolism ; Transcription Factors/physiology ; bcl-2-Associated X Protein
Chemical Substances	BAX protein, human ; BNIP3 protein, human ; DNA-Binding Proteins ; HIF1A protein, human ; Hypoxia-Inducible Factor 1 ; Hypoxia-Inducible Factor 1, alpha Subunit ; Inhibitor of Apoptosis Proteins ; Membrane Proteins ; Nuclear Proteins ; Proteins ; Proto-Oncogene Proteins ; Proto-Oncogene Proteins c-bcl-2 ; Transcription Factors ; bcl-2-Associated X Protein
Language	English
Publishing date	2004-10-08
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	80261-x
ISSN	1472-4146 ; 0021-9746
ISSN (online)	1472-4146
ISSN	0021-9746
DOI	10.1136/jcp.2003.015032
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Article ; Online: Epstein-Barr virus transcription activator R upregulates BARF1 expression by direct binding to its promoter, independent of methylation.

Hoebe, E K / Wille, C / Hopmans, E S / Robinson, A R / Middeldorp, J M / Kenney, S C / Greijer, A E

Journal of virology

2012 Volume 86, Issue 20, Page(s) 11322–11332

Abstract: Epstein-Barr virus (EBV) BamHI-A rightward frame 1 (BARF1) is considered a major viral oncogene in epithelial cells and has immune-modulating properties. However, in B cells and lymphomas, BARF1 expression is restricted to the viral lytic replication ... ...

Abstract	Epstein-Barr virus (EBV) BamHI-A rightward frame 1 (BARF1) is considered a major viral oncogene in epithelial cells and has immune-modulating properties. However, in B cells and lymphomas, BARF1 expression is restricted to the viral lytic replication cycle. In this report, the transcriptional regulation of BARF1 during lytic replication is unraveled. Bisulfite sequencing of various cell lines indicated a high level of methylation of the BARF1 gene control region. A BARF1 promoter luciferase reporter construct was created using a CpG-free vector, enabling true assessment of promoter methylation. Induction of the EBV lytic cycle is mediated by the immediate-early proteins BZLF1 (Z) and BRLF1 (R). R was found to activate expression of the BARF1 promoter up to 250-fold independently of Z and unaffected by BARF1 promoter methylation. Chromatin immunoprecipitation (ChIP), electrophoretic mobility shift assay (EMSA), and specific mutagenesis of the R-responsive elements (RREs) demonstrated direct binding of R to RREs between nucleotides -554 and -327 relative to the BARF1 transcriptional ATG start site. The kinetics of BARF1 expression upon transactivation by R showed that BARF1 mRNA was expressed within 6 h in the context of the viral genome. In conclusion, expression of the BARF1 protein during lytic replication is regulated by direct binding of R to multiple RREs in the gene control region and is independent of the promoter methylation status. The early kinetics of BARF1 upon transactivation by R confirm its status as an early gene and emphasize the necessity of early immune modulation during lytic reactivation.
MeSH term(s)	Cell Line, Tumor ; Chromatin Immunoprecipitation ; Electrophoretic Mobility Shift Assay ; Gene Expression Regulation, Viral ; Genes, Viral ; Herpesvirus 4, Human/metabolism ; Humans ; Immediate-Early Proteins/genetics ; Immediate-Early Proteins/metabolism ; Methylation ; Mutation ; Promoter Regions, Genetic ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; RNA, Viral/biosynthesis ; Response Elements ; Trans-Activators/genetics ; Trans-Activators/metabolism ; Transcription Factors/genetics ; Transcription Factors/metabolism ; Viral Proteins/biosynthesis ; Viral Proteins/genetics
Chemical Substances	BARF1 protein, Human herpesvirus 4 ; BRLF1 protein, Human herpesvirus 4 ; BZLF1 protein, Herpesvirus 4, Human ; Immediate-Early Proteins ; RNA, Messenger ; RNA, Viral ; Trans-Activators ; Transcription Factors ; Viral Proteins
Language	English
Publishing date	2012-08-15
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80174-4
ISSN	1098-5514 ; 0022-538X
ISSN (online)	1098-5514
ISSN	0022-538X
DOI	10.1128/JVI.01161-12
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In stock of ZB MED Cologne/Königswinter

Order via subito