Cell Cycle Independent Role of Cyclin D3 in Host Restriction of Influenza Virus Infection

Ying Fan, Chris Ka-Pun Mok, Michael Chi Wai Chan, Yang Zhang, Béatrice Nal-Rogier, François Kien, Roberto Bruzzone, Sumana Sanyal (Lead / Corresponding author)

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Abstract

To identify new host factors that modulate the replication of influenza A virus, we performed a yeast two-hybrid screen using the cytoplasmic tail of matrix protein 2 from the highly pathogenic H5N1 strain. The screen revealed a high-score interaction with cyclin D3, a key regulator of cell cycle early G1 phase. M2-cyclin D3 interaction was validated through GST pull-down and recapitulated in influenza A/WSN/33-infected cells. Knockdown of Ccnd3 by small interfering RNA significantly enhanced virus progeny titers in cell culture supernatants. Interestingly, the increase in virus production was due to cyclin D3 deficiency per se, and not merely a consequence of cell cycle deregulation. A combined knockdown of Ccnd3 and Rb1, which rescued cell cycle progression into the S phase, failed to normalize virus production. Infection by IAV triggered redistribution of cyclin D3 from the nucleus to the cytoplasm followed by its proteasomal degradation. When over-expressed in HEK 293T cells cyclin D3 impaired binding of M2 with M1, which is essential for proper assembly of progeny virions, lending further support to its role as a putative restriction factor. Our study describes the identification and characterization of cyclin D3 as a novel interactor of influenza A virus M2 protein. We hypothesize that competitive inhibition of M1-M2 interaction by cyclin D3 impairs infectious virion formation and results in attenuated virus production. In addition, we provide mechanistic insights into the dynamic interplay of influenza virus with the host cell cycle machinery during infection.

Original languageEnglish
Pages (from-to)5070-5088
Number of pages19
JournalJournal of Biological Chemistry
Volume292
Issue number12
Early online date27 Jan 2017
DOIs
Publication statusPublished - 24 Mar 2017

Fingerprint

Cyclin D3
Virus Diseases
Orthomyxoviridae
Viruses
Cell Cycle
Cells
Virion
Deregulation
HEK293 Cells
Influenza A virus
G1 Phase
Infection
Viral Load
S Phase
Cell culture
Yeast
Human Influenza
Small Interfering RNA
Machinery
Cytoplasm

Keywords

  • Cyclin D3
  • M2
  • Restriction factor
  • Cell cycle arrest
  • Influenza

Cite this

Fan, Y., Mok, C. K-P., Chan, M. C. W., Zhang, Y., Nal-Rogier, B., Kien, F., ... Sanyal, S. (2017). Cell Cycle Independent Role of Cyclin D3 in Host Restriction of Influenza Virus Infection. Journal of Biological Chemistry, 292(12), 5070-5088. https://doi.org/10.1074/jbc.M117.776112
Fan, Ying ; Mok, Chris Ka-Pun ; Chan, Michael Chi Wai ; Zhang, Yang ; Nal-Rogier, Béatrice ; Kien, François ; Bruzzone, Roberto ; Sanyal, Sumana. / Cell Cycle Independent Role of Cyclin D3 in Host Restriction of Influenza Virus Infection. In: Journal of Biological Chemistry. 2017 ; Vol. 292, No. 12. pp. 5070-5088.
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abstract = "To identify new host factors that modulate the replication of influenza A virus, we performed a yeast two-hybrid screen using the cytoplasmic tail of matrix protein 2 from the highly pathogenic H5N1 strain. The screen revealed a high-score interaction with cyclin D3, a key regulator of cell cycle early G1 phase. M2-cyclin D3 interaction was validated through GST pull-down and recapitulated in influenza A/WSN/33-infected cells. Knockdown of Ccnd3 by small interfering RNA significantly enhanced virus progeny titers in cell culture supernatants. Interestingly, the increase in virus production was due to cyclin D3 deficiency per se, and not merely a consequence of cell cycle deregulation. A combined knockdown of Ccnd3 and Rb1, which rescued cell cycle progression into the S phase, failed to normalize virus production. Infection by IAV triggered redistribution of cyclin D3 from the nucleus to the cytoplasm followed by its proteasomal degradation. When over-expressed in HEK 293T cells cyclin D3 impaired binding of M2 with M1, which is essential for proper assembly of progeny virions, lending further support to its role as a putative restriction factor. Our study describes the identification and characterization of cyclin D3 as a novel interactor of influenza A virus M2 protein. We hypothesize that competitive inhibition of M1-M2 interaction by cyclin D3 impairs infectious virion formation and results in attenuated virus production. In addition, we provide mechanistic insights into the dynamic interplay of influenza virus with the host cell cycle machinery during infection.",
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Fan, Y, Mok, CK-P, Chan, MCW, Zhang, Y, Nal-Rogier, B, Kien, F, Bruzzone, R & Sanyal, S 2017, 'Cell Cycle Independent Role of Cyclin D3 in Host Restriction of Influenza Virus Infection', Journal of Biological Chemistry, vol. 292, no. 12, pp. 5070-5088. https://doi.org/10.1074/jbc.M117.776112

Cell Cycle Independent Role of Cyclin D3 in Host Restriction of Influenza Virus Infection. / Fan, Ying; Mok, Chris Ka-Pun; Chan, Michael Chi Wai; Zhang, Yang; Nal-Rogier, Béatrice; Kien, François; Bruzzone, Roberto; Sanyal, Sumana (Lead / Corresponding author).

In: Journal of Biological Chemistry, Vol. 292, No. 12, 24.03.2017, p. 5070-5088.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Cell Cycle Independent Role of Cyclin D3 in Host Restriction of Influenza Virus Infection

AU - Fan, Ying

AU - Mok, Chris Ka-Pun

AU - Chan, Michael Chi Wai

AU - Zhang, Yang

AU - Nal-Rogier, Béatrice

AU - Kien, François

AU - Bruzzone, Roberto

AU - Sanyal, Sumana

N1 - This work was supported by the Research Fund for the Control of Infectious Diseases (Grant RFCID 11101332) and the Area of Excellence Scheme of the University Grants Committee (Grant AoE/M-12/06) and partially supported by grant from the Research Grants Council of the Hong Kong Special Administrative Region, China (Project No. T11-705/14N).

PY - 2017/3/24

Y1 - 2017/3/24

N2 - To identify new host factors that modulate the replication of influenza A virus, we performed a yeast two-hybrid screen using the cytoplasmic tail of matrix protein 2 from the highly pathogenic H5N1 strain. The screen revealed a high-score interaction with cyclin D3, a key regulator of cell cycle early G1 phase. M2-cyclin D3 interaction was validated through GST pull-down and recapitulated in influenza A/WSN/33-infected cells. Knockdown of Ccnd3 by small interfering RNA significantly enhanced virus progeny titers in cell culture supernatants. Interestingly, the increase in virus production was due to cyclin D3 deficiency per se, and not merely a consequence of cell cycle deregulation. A combined knockdown of Ccnd3 and Rb1, which rescued cell cycle progression into the S phase, failed to normalize virus production. Infection by IAV triggered redistribution of cyclin D3 from the nucleus to the cytoplasm followed by its proteasomal degradation. When over-expressed in HEK 293T cells cyclin D3 impaired binding of M2 with M1, which is essential for proper assembly of progeny virions, lending further support to its role as a putative restriction factor. Our study describes the identification and characterization of cyclin D3 as a novel interactor of influenza A virus M2 protein. We hypothesize that competitive inhibition of M1-M2 interaction by cyclin D3 impairs infectious virion formation and results in attenuated virus production. In addition, we provide mechanistic insights into the dynamic interplay of influenza virus with the host cell cycle machinery during infection.

AB - To identify new host factors that modulate the replication of influenza A virus, we performed a yeast two-hybrid screen using the cytoplasmic tail of matrix protein 2 from the highly pathogenic H5N1 strain. The screen revealed a high-score interaction with cyclin D3, a key regulator of cell cycle early G1 phase. M2-cyclin D3 interaction was validated through GST pull-down and recapitulated in influenza A/WSN/33-infected cells. Knockdown of Ccnd3 by small interfering RNA significantly enhanced virus progeny titers in cell culture supernatants. Interestingly, the increase in virus production was due to cyclin D3 deficiency per se, and not merely a consequence of cell cycle deregulation. A combined knockdown of Ccnd3 and Rb1, which rescued cell cycle progression into the S phase, failed to normalize virus production. Infection by IAV triggered redistribution of cyclin D3 from the nucleus to the cytoplasm followed by its proteasomal degradation. When over-expressed in HEK 293T cells cyclin D3 impaired binding of M2 with M1, which is essential for proper assembly of progeny virions, lending further support to its role as a putative restriction factor. Our study describes the identification and characterization of cyclin D3 as a novel interactor of influenza A virus M2 protein. We hypothesize that competitive inhibition of M1-M2 interaction by cyclin D3 impairs infectious virion formation and results in attenuated virus production. In addition, we provide mechanistic insights into the dynamic interplay of influenza virus with the host cell cycle machinery during infection.

KW - Cyclin D3

KW - M2

KW - Restriction factor

KW - Cell cycle arrest

KW - Influenza

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