High resolution cryo-EM structure of the helical RNA-bound Hantaan virus nucleocapsid reveals its assembly mechanisms

Arragain, Benoît; Reguera, Juan; Desfosses, Ambroise; Gutsche, Irina; Schoehn, Guy; Malet, Hélène

doi:10.7554/elife.43075

Cited by 31 publications

(37 citation statements)

References 35 publications

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“…At all the time points examined, these NP assemblies closely co-localized with TULV RNA, regardless of its negative or positive sense polarity ( Figure 6). This close association between all hantavirus RNA and NP is consistent with the known structure of the hantavirus RNP segments, in which hantaviral vRNAs and cRNAs are always found in close association with NP, forming extended helical assemblies [31]. Interestingly, our observations also suggest that TULV mRNAs, which will likely comprise the majority of positive sense TULV-specific RNA in the infected cell [30], also remain in close proximity with NP.…”

Section: Discussionsupporting

confidence: 87%

The RNA Replication Site of Tula Orthohantavirus Resides within a Remodelled Golgi Network

Davies

Chadwick

Hewson

et al. 2020

Cells

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The family Hantaviridae within the Bunyavirales order comprises tri-segmented negative sense RNA viruses, many of which are rodent-borne emerging pathogens associated with fatal human disease. In contrast, hantavirus infection of corresponding rodent hosts results in inapparent or latent infections, which can be recapitulated in cultured cells that become persistently infected. In this study, we used Tula virus (TULV) to investigate the location of hantavirus replication during early, peak and persistent phases of infection, over a 30-day time course. Using immunofluorescent (IF) microscopy, we showed that the TULV nucleocapsid protein (NP) is distributed within both punctate and filamentous structures, with the latter increasing in size as the infection progresses. Transmission electron microscopy of TULV-infected cell sections revealed these filamentous structures comprised aligned clusters of filament bundles. The filamentous NP-associated structures increasingly co-localized with the Golgi and with the stress granule marker TIA-1 over the infection time course, suggesting a redistribution of these cellular organelles. The analysis of the intracellular distribution of TULV RNAs using fluorescent in-situ hybridization revealed that both genomic and mRNAs co-localized with Golgi-associated filamentous compartments that were positive for TIA. These results show that TULV induces a dramatic reorganization of the intracellular environment, including the establishment of TULV RNA synthesis factories in re-modelled Golgi compartments.

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Section: Discussionsupporting

confidence: 87%

The RNA Replication Site of Tula Orthohantavirus Resides within a Remodelled Golgi Network

Davies

Chadwick

Hewson

et al. 2020

Cells

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“…The investigations of a possible cap-binding function of Junin, Tacaribe, and Pichinde arenavirus N proteins did not provide sufficient proof for a direct interaction of N with cap-structures and, additionally, important experimental controls were not presented [35]. In the crystal structure of hantavirus N protein as well as in the cryo-electron microscopy structure of hantavirus nucleocapsid, no apparent cap-binding site could be observed [38,39]. The studies suggesting a cap-binding site in hantavirus N protein did neither include sufficient evidence nor include mutational analyses to validate this hypothesis [31][32][33].…”

Section: The Cap-binding Functionmentioning

confidence: 98%

The Cap-Snatching Mechanism of Bunyaviruses

Olschewski

Cusack

Rosenthal

2020

Trends in Microbiology

110

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In common with all segmented negative-sense RNA viruses, bunyavirus transcripts contain heterologous sequences at their 5′ termini originating from capped host cell RNAs. These heterologous sequences are acquired by a socalled cap-snatching mechanism. Whereas for nuclear replicating influenza virus the source of capped primers as well as the cap-binding and endonuclease activities of the viral polymerase needed for cap snatching have been functionally and structurally well characterized, our knowledge on the expected counterparts of cytoplasmic replicating bunyaviruses is still limited and controversial. This review focuses on the cap-snatching mechanism of bunyaviruses in the light of recent structural and functional data. Bunyavirus Transcription and Genome Replication The order of Bunyavirales, which was established in 2018, accommodates a diverse group of viruses formerly separated into the Arenaviridae and Bunyaviridae families [1]. The Bunyavirales order contains important human pathogens such as Lassa virus (LASV; family Arenaviridae), Crimean-Congo hemorrhagic fever virus (family Nairoviridae), Rift Valley fever virus (RVFV, family Phenuiviridae), Hantaan virus (family Hantaviridae), and La Crosse virus (family Peribunyaviridae). Highlights Endonuclease domains have been located in many different bunyavirus L proteins and can be classified as His+ or His-metal-dependent endonucleases. An active cap-binding domain (CBD) has been identified in the C-terminal region of bunyavirus L protein. Despite very low sequence identity, this domain is very similar to influenza virus PB2 CBD on the structural level. Atomic structures of bunyavirus N protein do not provide evidence for the hypothesized N protein cap-binding site. It remains unclear whether a functionally relevant cap-specific binding site exists in any bunyavirus N protein. The comparably low affinity of bunyavirus CBD for cap-structures indicates the necessity for further regions of the L protein or other viral or cellular proteins to be involved in cap binding.

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“…In Hantaan virus, there are three nucleotides per subunit. The cross subunit interactions are much more extensive than nucleocapsids from other viruses [17]. Each subunit has domain interchanges with +3 and -3 subunits, including residues in termini and five protruding loops.…”

Section: Assembly Of the Nucleocapsidmentioning

confidence: 99%

Nucleocapsid Structure of Negative Strand RNA Virus

Luo

Terrell

Mcmanus

2020

Viruses

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Negative strand RNA viruses (NSVs) include many important human pathogens, such as influenza virus, Ebola virus, and rabies virus. One of the unique characteristics that NSVs share is the assembly of the nucleocapsid and its role in viral RNA synthesis. In NSVs, the single strand RNA genome is encapsidated in the linear nucleocapsid throughout the viral replication cycle. Subunits of the nucleocapsid protein are parallelly aligned along the RNA genome that is sandwiched between two domains composed of conserved helix motifs. The viral RNA-dependent-RNA polymerase (vRdRp) must recognize the protein–RNA complex of the nucleocapsid and unveil the protected genomic RNA in order to initiate viral RNA synthesis. In addition, vRdRp must continuously translocate along the protein–RNA complex during elongation in viral RNA synthesis. This unique mechanism of viral RNA synthesis suggests that the nucleocapsid may play a regulatory role during NSV replication.

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High resolution cryo-EM structure of the helical RNA-bound Hantaan virus nucleocapsid reveals its assembly mechanisms

Cited by 31 publications

References 35 publications

The RNA Replication Site of Tula Orthohantavirus Resides within a Remodelled Golgi Network

The RNA Replication Site of Tula Orthohantavirus Resides within a Remodelled Golgi Network

The Cap-Snatching Mechanism of Bunyaviruses

Nucleocapsid Structure of Negative Strand RNA Virus

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