Epstein-Barr virus BKRF4 gene product is required for efficient progeny production

H. M.Abdullah Al Masud, Takahiro Watanabe, Masahiro Yoshida, Yoshitaka Sato, Fumi Goshima, Hiroshi Kimura, Takayuki Murata

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Epstein-Barr virus (EBV), a member of human gammaherpesvirus, infects mainly B cells. EBV has two alternative life cycles, latent and lytic, and is reactivated occasionally from the latent stage to the lytic cycle. To combat EBV-associated disorders, understanding the molecular mechanisms of the EBV lytic replication cycle is also important. Here, we focused on an EBV lytic gene, BKRF4. Using our anti-BKRF4 antibody, we revealed that the BKRF4 gene product is expressed during the lytic cycle with late kinetics. To characterize the role of BKRF4, we constructed BKRF4- knockout mutants using the bacterial artificial chromosome (BAC) and CRISPR/Cas9 systems. Although disruption of the BKRF4 gene had almost no effect on viral protein expression and DNA synthesis, it significantly decreased progeny virion levels in HEK293 and Akata cells. Furthermore, we show that BKRF4 is involved not only in production of progeny virions but also in increasing the infectivity of the virus particles. Immunoprecipitation assays revealed that BKRF4 interacted with a virion protein, BGLF2. We showed that the C-terminal region of BKRF4 was critical for this interaction and for efficient progeny production. Immunofluorescence analysis revealed that BKRF4 partially colocalized with BGLF2 in the nucleus and perinuclear region. Finally, we showed that BKRF4 is a phosphorylated, possible tegument protein and that the EBV protein kinase BGLF4 may be important for this phosphorylation. Taken together, our data suggest that BKRF4 is involved in the production of infectious virions.

Original languageEnglish
Article numbere00975-17
JournalJournal of Virology
Volume91
Issue number23
DOIs
Publication statusPublished - 01-12-2017

Fingerprint

Human herpesvirus 4
Human Herpesvirus 4
virion
Virion
Genes
genes
Clustered Regularly Interspaced Short Palindromic Repeats
Bacterial Artificial Chromosomes
bacterial artificial chromosomes
knockout mutants
HEK293 Cells
viral proteins
Viral DNA
Viral Proteins
gene targeting
Virus Replication
Life Cycle Stages
Immunoprecipitation
protein kinases
Protein Kinases

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Masud, H. M. A. A., Watanabe, T., Yoshida, M., Sato, Y., Goshima, F., Kimura, H., & Murata, T. (2017). Epstein-Barr virus BKRF4 gene product is required for efficient progeny production. Journal of Virology, 91(23), [e00975-17]. https://doi.org/10.1128/JVI.00975-17
Masud, H. M.Abdullah Al ; Watanabe, Takahiro ; Yoshida, Masahiro ; Sato, Yoshitaka ; Goshima, Fumi ; Kimura, Hiroshi ; Murata, Takayuki. / Epstein-Barr virus BKRF4 gene product is required for efficient progeny production. In: Journal of Virology. 2017 ; Vol. 91, No. 23.
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abstract = "Epstein-Barr virus (EBV), a member of human gammaherpesvirus, infects mainly B cells. EBV has two alternative life cycles, latent and lytic, and is reactivated occasionally from the latent stage to the lytic cycle. To combat EBV-associated disorders, understanding the molecular mechanisms of the EBV lytic replication cycle is also important. Here, we focused on an EBV lytic gene, BKRF4. Using our anti-BKRF4 antibody, we revealed that the BKRF4 gene product is expressed during the lytic cycle with late kinetics. To characterize the role of BKRF4, we constructed BKRF4- knockout mutants using the bacterial artificial chromosome (BAC) and CRISPR/Cas9 systems. Although disruption of the BKRF4 gene had almost no effect on viral protein expression and DNA synthesis, it significantly decreased progeny virion levels in HEK293 and Akata cells. Furthermore, we show that BKRF4 is involved not only in production of progeny virions but also in increasing the infectivity of the virus particles. Immunoprecipitation assays revealed that BKRF4 interacted with a virion protein, BGLF2. We showed that the C-terminal region of BKRF4 was critical for this interaction and for efficient progeny production. Immunofluorescence analysis revealed that BKRF4 partially colocalized with BGLF2 in the nucleus and perinuclear region. Finally, we showed that BKRF4 is a phosphorylated, possible tegument protein and that the EBV protein kinase BGLF4 may be important for this phosphorylation. Taken together, our data suggest that BKRF4 is involved in the production of infectious virions.",
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Masud, HMAA, Watanabe, T, Yoshida, M, Sato, Y, Goshima, F, Kimura, H & Murata, T 2017, 'Epstein-Barr virus BKRF4 gene product is required for efficient progeny production', Journal of Virology, vol. 91, no. 23, e00975-17. https://doi.org/10.1128/JVI.00975-17

Epstein-Barr virus BKRF4 gene product is required for efficient progeny production. / Masud, H. M.Abdullah Al; Watanabe, Takahiro; Yoshida, Masahiro; Sato, Yoshitaka; Goshima, Fumi; Kimura, Hiroshi; Murata, Takayuki.

In: Journal of Virology, Vol. 91, No. 23, e00975-17, 01.12.2017.

Research output: Contribution to journalArticle

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Masud HMAA, Watanabe T, Yoshida M, Sato Y, Goshima F, Kimura H et al. Epstein-Barr virus BKRF4 gene product is required for efficient progeny production. Journal of Virology. 2017 Dec 1;91(23). e00975-17. https://doi.org/10.1128/JVI.00975-17