Structural insight into the essential PB1–PB2 subunit contact of the influenza virus RNA polymerase

Sugiyama, Kanako; Obayashi, E.; Kawaguchi, Akiko; Suzuki, Yukari; Tame, J.R.H.; Nagata, Kyosuke; Park, Sam-Yong

doi:10.1038/emboj.2009.138

Cited by 174 publications

(208 citation statements)

References 45 publications

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“…2). Some of these interactions have been verified by co-crystallisation [55][56][57] and functional studies. 58 Also, some additional PB2-PA and PB1-PB2 interactions have been described in reference 40 and 59, that may play structural and/or regulatory roles in the activity of the viral polymerase.…”

Section: The Polymerase Complexmentioning

confidence: 77%

The influenza virus RNA synthesis machine

et al. 2011

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Section: The Polymerase Complexmentioning

confidence: 77%

The influenza virus RNA synthesis machine

et al. 2011

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“…Each RdRp subunit is thought to play a distinct and essential role within the polymerase. Although extensive functional analyses have elucidated many aspects of the overall RdRp structure, little is known regarding control of RNA synthesis (3)(4)(5)(6)(7)(8)(9).…”

mentioning

confidence: 99%

Influenza A virus-generated small RNAs regulate the switch from transcription to replication

Perez

Varble

Sachidanandam³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

194

227

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The discovery of regulatory small RNAs continues to reshape paradigms in both molecular biology and virology. Here we describe examples of influenza A virus-derived small viral RNAs (svRNAs). svRNAs are 22-27 nt in length and correspond to the 5′ end of each of the viral genomic RNA (vRNA) segments. Expression of svRNA correlates with the accumulation of vRNA and a bias in RNAdependent RNA polymerase (RdRp) activity from transcription toward genome replication. Synthesis of svRNA requires the RdRp, nucleoprotein and the nuclear export protein NS2. In addition, svRNA is detectable during replication of various influenza A virus subtypes across multiple host species and associates physically with the RdRp. We demonstrate that depletion of svRNA has a minimal impact on mRNA and complementary vRNA (cRNA) but results in a dramatic loss of vRNA in a segment-specific manner. We propose that svRNA triggers the viral switch from transcription to replication through interactions with the viral polymerase machinery. Taken together, the discovery of svRNA redefines the mechanistic switch of influenza virus transcription/replication and provides a potential target for broad-range, anti-influenza virus-based therapeutics.microRNA | replicase | replication switch | RNA dependent RNA polymerase | transcriptase

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“…Many high-yield IVSs produced by natural reassortment contained the PB1 segment of the circulating strain rather than the PB1 PR8 segment (Sugiyama et al, 2009), but there are also examples in which the indigenous PB1 of the circulating strain was not beneficial for producing a more efficiently replicating IVS (Arranz et al, 2012;Hemerka et al, 2009). Therefore, it would be highly desirable to identify a 'generic' (broadly or even universally applicable) PB1 segment that may replace the PB1 PR8 segment to overcome possible restrictions of replication observed for many 6+2 IVSs (Abt et al, 2011;Sugiyama et al, 2009;Wanitchang et al, 2010). Here, we explored the capability of the PB1 Gi segment to support the replication of different recombinant PR8-based 5+2+1 IVSs.…”

Section: Discussionmentioning

confidence: 99%

The PB1 segment of an influenza A virus H1N1 2009pdm isolate enhances the replication efficiency of specific influenza vaccine strains in cell culture and embryonated eggs

Mostafa

Kanrai

Ziebuhr

et al. 2016

Journal of General Virology

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Influenza vaccine strains (IVSs) contain the haemagglutinin (HA) and neuraminidase (NA) genome segments of relevant circulating strains in the genetic background of influenza A/ PR/8/1934 virus (PR8). Previous work has shown that the nature of the PB1 segment may be a limiting factor for the efficient production of IVSs. Here, we showed that the PB1 segment (PB1 Gi ) from the 2009 pandemic influenza A virus (IAV) A/Giessen/06/2009 (Gi wt, H1N1pdm) may help to resolve (some of) these limitations. We produced a set of recombinant PR8-derived viruses that contained (i) the HA and NA segments from representative IAV strains (H3N2, H5N1, H7N9, H9N2); (ii) the PB1 segment from PR8 or Gi wt, respectively; and (iii) the remaining five genome segments from PR8. Viruses containing the PB1 Gi segment, together with the heterologous HA/NA segments and five PR8 segments (5+2+1), replicated to higher titres compared with their 6+2 counterparts containing six PR8 segments and the equivalent heterologous HA/NA segments. Compared with PB1 PR8 -containing IVSs, viruses with the PB1 Gi segment replicated to higher or similar titres in both cell culture and embryonated eggs, most profoundly IVSs of the H5N1 and H7N9 subtype, which are known to grow poorly in these systems. IVSs containing either the PB1 Gi or the cognate PB1 segment of the respective specific HA/NA donor strain showed enhanced or similar virus replication levels. This study suggests that substitution of PB1 PR8 with the PB1 Gi segment may greatly improve the large-scale production of PR8-derived IVSs, especially of those known to replicate poorly in vitro. INTRODUCTIONInfluenza viruses belong to the family Orthomyxoviridae and are classified into three genera termed Influenza virus A, B and C. Among these, influenza A viruses (IAVs) pose the greatest threat to human and animal health. They contain eight segments of negative-sense ssRNA (Ritchey et al., 1976;Shaw & Palese, 2013). The eight genomic segments encode at least 10 viral proteins, comprising the three subunits of the viral RNA polymerase complex [polymerase basic protein 2 (PB2), polymerase basic protein 1 (PB1) and polymerase acidic protein (PA)], the surface glycoproteins [haemagglutinin (HA) and neuraminidase (NA)], the nucleoprotein (NP), the matrix protein (M1), the ion channel protein (M2), the non-structural protein 1 (NS1) and the nuclear export protein (NEP). IAVs are genetically diverse, with new strains rapidly emerging by point mutations introduced by the low-fidelity viral RNA replicase (potentially resulting in antigenic drift if changes occur in the HA and/or NA coding sequences and affect important antigenic epitopes) or by genetic reassortment of genomic segments (potentially leading to antigenic shift if antigenically distant HA and NA segments are incorporated). Minor and major antigenic changes in the HA and NA proteins of circulating IAV are important driving forces for the emergence of new strains causing epidemics or even major pandemics (Neumann et al., 2009;Peng et al., 2014;Smith...

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Structural insight into the essential PB1–PB2 subunit contact of the influenza virus RNA polymerase

Cited by 174 publications

References 45 publications

The influenza virus RNA synthesis machine

The influenza virus RNA synthesis machine

Influenza A virus-generated small RNAs regulate the switch from transcription to replication

The PB1 segment of an influenza A virus H1N1 2009pdm isolate enhances the replication efficiency of specific influenza vaccine strains in cell culture and embryonated eggs

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