The RNA Polymerase PB2 Subunit of Influenza A/HongKong/156/1997 (H5N1) Restrict the Replication of Reassortant Ribonucleoprotein Complexes

Nakazono, Yoko; Hara, Koyu; Kashiwagi, Takanari; Hamada, Nobuyuki; Watanabe, Hiroshi

doi:10.1371/journal.pone.0032634

Cited by 13 publications

(17 citation statements)

References 48 publications

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“…However, notable differences in the profile of transcriptive (mRNA) and replicative (vRNA and cRNA) intermediates were observed between 33°C and 37°C. Similar to earlier study, the K627E mutation significantly reduced vRNA and cRNA promoter binding activities of PB2 in avian H5N1 virus [35]. At 37°C, all three mutations only resulted in the elevation of vRNA level but not mRNA and cRNA levels, suggesting that these mutations may only enhance the polymerase activity of cRNA-dependent vRNA synthesis but not the activity of vRNA-dependent mRNA and cRNA synthesis.…”

Section: Discussionsupporting

confidence: 87%

Replication and Transcription Activities of Ribonucleoprotein Complexes Reconstituted from Avian H5N1, H1N1pdm09 and H3N2 Influenza A Viruses

Ngai

Chan

2013

PLoS ONE

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Avian influenza viruses pose a serious pandemic threat to humans. Better knowledge on cross-species adaptation is important. This study examined the replication and transcription efficiency of ribonucleoprotein complexes reconstituted by plasmid co-transfection between H5N1, H1N1pdm09 and H3N2 influenza A viruses, and to identify mutations in the RNA polymerase subunit that affect human adaptation. Viral RNA polymerase subunits PB1, PB2, PA and NP derived from influenza viruses were co-expressed with pPolI-vNP-Luc in human cells, and with its function evaluated by luciferase reporter assay. A quantitative RT-PCR was used to measure vRNA, cRNA, and mRNA levels for assessing the replication and transcription efficiency. Mutations in polymerase subunit were created to identify signature of increased human adaptability. H5N1 ribonucleoprotein complexes incorporated with PB2 derived from H1N1pdm09 and H3N2 viruses increased the polymerase activity in human cells. Furthermore, single amino acid substitutions at PB2 of H5N1 could affect polymerase activity in a temperature-dependent manner. By using a highly sensitive quantitative reverse transcription-polymerase chain reaction, an obvious enhancement in replication and transcription activities of ribonucleoproteins was observed by the introduction of lysine at residue 627 in the H5N1 PB2 subunit. Although less strongly in polymerase activity, E158G mutation appeared to alter the accumulation of H5N1 RNA levels in a temperature-dependent manner, suggesting a temperature-dependent mechanism in regulating transcription and replication exists. H5N1 viruses can adapt to humans either by acquisition of PB2 from circulating human-adapted viruses through reassortment, or by mutations at critical sites in PB2. This information may help to predict the pandemic potential of newly emerged influenza strains, and provide a scientific basis for stepping up surveillance measures and vaccine production.

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Section: Discussionsupporting

confidence: 87%

Replication and Transcription Activities of Ribonucleoprotein Complexes Reconstituted from Avian H5N1, H1N1pdm09 and H3N2 Influenza A Viruses

Ngai

Chan

2013

PLoS ONE

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“…It has been reported that amino acid changes in PB2, such as T271A, K627E, and D701N, are related to adaptation of avian influenza virus to humans and virulence (15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25). In this study, we found that the residue at position 339 in the cap-binding pocket displays a significant and widespread amino acid polymorphism.…”

mentioning

confidence: 48%

Structural and Functional Characterization of K339T Substitution Identified in the PB2 Subunit Cap-binding Pocket of Influenza A Virus

Liu

Qin

Meng

et al. 2013

Journal of Biological Chemistry

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Background: Amino acid changes in PB2 are associated with evolution of influenza virus. Results: K339T substitution in PB2 cap reduces the cap binding affinity, polymerase activity, RNA synthesis activity, and murine mortality. Conclusion: Substitution in PB2 cap modulates the polymerase activity and virulence by regulating the cap binding activity. Significance: We identified and characterized an emerging K339T substitution in PB2 cap .

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“…was observed with full-length PB2 627K (129,130), leaving the role of the 627 domain in RNA binding unresolved.…”

Section: Figmentioning

confidence: 99%

Adaptation of Avian Influenza A Virus Polymerase in Mammals To Overcome the Host Species Barrier

Mänz

Schwemmle

Brunotte

2013

J Virol

197

186

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Avian influenza A viruses, such as the highly pathogenic avian H5N1 viruses, sporadically enter the human population but often do not transmit between individuals. In rare cases, however, they establish a new lineage in humans. In addition to well-characterized barriers to cell entry, one major hurdle which avian viruses must overcome is their poor polymerase activity in human cells. There is compelling evidence that these viruses overcome this obstacle by acquiring adaptive mutations in the polymerase subunits PB1, PB2, and PA and the nucleoprotein (NP) as well as in the novel polymerase cofactor nuclear export protein (NEP). Recent findings suggest that synthesis of the viral genome may represent the major defect of avian polymerases in human cells. While the precise mechanisms remain to be unveiled, it appears that a broad spectrum of polymerase adaptive mutations can act collectively to overcome this defect. Thus, identification and monitoring of emerging adaptive mutations that further increase polymerase activity in human cells are critical to estimate the pandemic potential of avian viruses.

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The RNA Polymerase PB2 Subunit of Influenza A/HongKong/156/1997 (H5N1) Restrict the Replication of Reassortant Ribonucleoprotein Complexes

Cited by 13 publications

References 48 publications

Replication and Transcription Activities of Ribonucleoprotein Complexes Reconstituted from Avian H5N1, H1N1pdm09 and H3N2 Influenza A Viruses

Replication and Transcription Activities of Ribonucleoprotein Complexes Reconstituted from Avian H5N1, H1N1pdm09 and H3N2 Influenza A Viruses

Structural and Functional Characterization of K339T Substitution Identified in the PB2 Subunit Cap-binding Pocket of Influenza A Virus

Adaptation of Avian Influenza A Virus Polymerase in Mammals To Overcome the Host Species Barrier

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