Epstein-Barr virus polymerase processivity factor enhances BALF2 promoter transcription as a coactivator for the BZLF1immediate-early protein

Sanae Nakayama, Takayuki Murata, Kazutaka Murayama, Yoshihiro Yasui, Yoshitaka Sato, Ayumi Kudoh, Satoko Iwahori, Hiroki Isomura, Teru Kanda, Tatsuya Tsurumi

Research output: Contribution to journalArticle

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Abstract

The Epstein-Barr virus (EBV) BMRF1 protein is an essential replication protein acting at viral replication forks as a viral DNA polymerase processivity factor, whereas the BALF2 protein is a singles-tranded DNA-binding protein that also acts at replication forks and is most abundantly expressed during viral productive replication. Here we document that the BMRF1 protein evidently enhances viral BZLF1 transcription factor-mediated transactivation of the BALF2 gene promoter. Mutagenesis and electrophoretic mobility shift assays demonstrated the BALF2 promoter to harbor two BZLF1 protein-binding sites (BZLF1-responsive elements). Direct binding of the BZLF1 protein to BZLF1-responsive elements and physical interaction between BZLF1 and BMRF1 proteins are prerequisite for the BMRF1 protein up-regulation of the BALF2 gene promoter. A monomeric mutant, C95E, which is defective in homodimerization, could still interact and enhance BZLF1-mediated transactivation. Furthermore although EBV protein kinase phosphorylates BMRF1 protein extensively, it turned out that phosphorylation of the protein by the kinase is inhibitory to the enhancement of the BZLF1-mediated transactivation of BALF2 promoter. Exogenous expression of BMRF1 protein augmented BALF2 expression in HEK293 cells harboring the EBV genome but lacking BMRF1 and BALF5 genes, demonstrating functions as a transcriptional regulator in the context of viral infection. Overall the BMRF1 protein is a multifunctional protein that cannot only act as a DNA polymerase processivity factor but also enhances BALF2 promoter transcription as a coactivator for the BZLF1 protein, regulating the expression level of viral single-stranded DNA-binding protein.

Original languageEnglish
Pages (from-to)21557-21568
Number of pages12
JournalJournal of Biological Chemistry
Volume284
Issue number32
DOIs
Publication statusPublished - 07-08-2009
Externally publishedYes

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Transcription
Human Herpesvirus 4
Viruses
Proteins
Genes
Transcriptional Activation
Viral DNA
DNA-Binding Proteins
DNA-Directed DNA Polymerase
Protein Kinases
Electrophoretic mobility
HEK293 Cells
Mutagenesis
Phosphorylation
Electrophoretic Mobility Shift Assay
Virus Diseases
Protein Binding
Ports and harbors
Carrier Proteins
Transcription Factors

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Nakayama, Sanae ; Murata, Takayuki ; Murayama, Kazutaka ; Yasui, Yoshihiro ; Sato, Yoshitaka ; Kudoh, Ayumi ; Iwahori, Satoko ; Isomura, Hiroki ; Kanda, Teru ; Tsurumi, Tatsuya. / Epstein-Barr virus polymerase processivity factor enhances BALF2 promoter transcription as a coactivator for the BZLF1immediate-early protein. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 32. pp. 21557-21568.
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abstract = "The Epstein-Barr virus (EBV) BMRF1 protein is an essential replication protein acting at viral replication forks as a viral DNA polymerase processivity factor, whereas the BALF2 protein is a singles-tranded DNA-binding protein that also acts at replication forks and is most abundantly expressed during viral productive replication. Here we document that the BMRF1 protein evidently enhances viral BZLF1 transcription factor-mediated transactivation of the BALF2 gene promoter. Mutagenesis and electrophoretic mobility shift assays demonstrated the BALF2 promoter to harbor two BZLF1 protein-binding sites (BZLF1-responsive elements). Direct binding of the BZLF1 protein to BZLF1-responsive elements and physical interaction between BZLF1 and BMRF1 proteins are prerequisite for the BMRF1 protein up-regulation of the BALF2 gene promoter. A monomeric mutant, C95E, which is defective in homodimerization, could still interact and enhance BZLF1-mediated transactivation. Furthermore although EBV protein kinase phosphorylates BMRF1 protein extensively, it turned out that phosphorylation of the protein by the kinase is inhibitory to the enhancement of the BZLF1-mediated transactivation of BALF2 promoter. Exogenous expression of BMRF1 protein augmented BALF2 expression in HEK293 cells harboring the EBV genome but lacking BMRF1 and BALF5 genes, demonstrating functions as a transcriptional regulator in the context of viral infection. Overall the BMRF1 protein is a multifunctional protein that cannot only act as a DNA polymerase processivity factor but also enhances BALF2 promoter transcription as a coactivator for the BZLF1 protein, regulating the expression level of viral single-stranded DNA-binding protein.",
author = "Sanae Nakayama and Takayuki Murata and Kazutaka Murayama and Yoshihiro Yasui and Yoshitaka Sato and Ayumi Kudoh and Satoko Iwahori and Hiroki Isomura and Teru Kanda and Tatsuya Tsurumi",
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Nakayama, S, Murata, T, Murayama, K, Yasui, Y, Sato, Y, Kudoh, A, Iwahori, S, Isomura, H, Kanda, T & Tsurumi, T 2009, 'Epstein-Barr virus polymerase processivity factor enhances BALF2 promoter transcription as a coactivator for the BZLF1immediate-early protein', Journal of Biological Chemistry, vol. 284, no. 32, pp. 21557-21568. https://doi.org/10.1074/jbc.M109.015685

Epstein-Barr virus polymerase processivity factor enhances BALF2 promoter transcription as a coactivator for the BZLF1immediate-early protein. / Nakayama, Sanae; Murata, Takayuki; Murayama, Kazutaka; Yasui, Yoshihiro; Sato, Yoshitaka; Kudoh, Ayumi; Iwahori, Satoko; Isomura, Hiroki; Kanda, Teru; Tsurumi, Tatsuya.

In: Journal of Biological Chemistry, Vol. 284, No. 32, 07.08.2009, p. 21557-21568.

Research output: Contribution to journalArticle

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T1 - Epstein-Barr virus polymerase processivity factor enhances BALF2 promoter transcription as a coactivator for the BZLF1immediate-early protein

AU - Nakayama, Sanae

AU - Murata, Takayuki

AU - Murayama, Kazutaka

AU - Yasui, Yoshihiro

AU - Sato, Yoshitaka

AU - Kudoh, Ayumi

AU - Iwahori, Satoko

AU - Isomura, Hiroki

AU - Kanda, Teru

AU - Tsurumi, Tatsuya

PY - 2009/8/7

Y1 - 2009/8/7

N2 - The Epstein-Barr virus (EBV) BMRF1 protein is an essential replication protein acting at viral replication forks as a viral DNA polymerase processivity factor, whereas the BALF2 protein is a singles-tranded DNA-binding protein that also acts at replication forks and is most abundantly expressed during viral productive replication. Here we document that the BMRF1 protein evidently enhances viral BZLF1 transcription factor-mediated transactivation of the BALF2 gene promoter. Mutagenesis and electrophoretic mobility shift assays demonstrated the BALF2 promoter to harbor two BZLF1 protein-binding sites (BZLF1-responsive elements). Direct binding of the BZLF1 protein to BZLF1-responsive elements and physical interaction between BZLF1 and BMRF1 proteins are prerequisite for the BMRF1 protein up-regulation of the BALF2 gene promoter. A monomeric mutant, C95E, which is defective in homodimerization, could still interact and enhance BZLF1-mediated transactivation. Furthermore although EBV protein kinase phosphorylates BMRF1 protein extensively, it turned out that phosphorylation of the protein by the kinase is inhibitory to the enhancement of the BZLF1-mediated transactivation of BALF2 promoter. Exogenous expression of BMRF1 protein augmented BALF2 expression in HEK293 cells harboring the EBV genome but lacking BMRF1 and BALF5 genes, demonstrating functions as a transcriptional regulator in the context of viral infection. Overall the BMRF1 protein is a multifunctional protein that cannot only act as a DNA polymerase processivity factor but also enhances BALF2 promoter transcription as a coactivator for the BZLF1 protein, regulating the expression level of viral single-stranded DNA-binding protein.

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