Architecture and regulation of negative-strand viral enzymatic machinery

Kranzusch, Philip J.; Whelan, Sean P. J.

doi:10.4161/rna.20345

Cited by 29 publications

(29 citation statements)

References 106 publications

(140 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Previous studies have suggested that bunyavirus NPs cannot well protect the bound RNA against exogenous RNases (1,5). These observations are consistent with the finding that the NPs of nonsegmented -ssRNA viruses can protect the bound RNA, in contrast to the NPs of segmented -ssRNA viruses, which cannot (9).…”

Section: Resultssupporting

confidence: 82%

“…Another interesting observation from crystallographic studies on all of the bunyavirus-encoded NPs is that all of them cannot properly protect the packaged RNA against exogenous RNases. This result is consistent with the finding that bound RNA can be well protected by NPs from nonsegmented -ssRNA viruses but not by NPs from segmented -ssRNA viruses (8,9). Bunyamwera virus (BUNV), which can cause severe hemorrhagic fever and other diseases in livestock and humans (10), is the prototypic member of both the Orthobunyavirus genus and the entire Bunyaviridae family.…”

supporting

confidence: 79%

See 1 more Smart Citation

Bunyamwera virus possesses a distinct nucleocapsid protein to facilitate genome encapsidation

Li¹,

Wang

Pan

et al. 2013

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

View full text Add to dashboard Cite

Bunyamwera virus (BUNV), which belongs to the genus Orthobunyavirus, is the prototypical virus of the Bunyaviridae family. Similar to other negative-sense single-stranded RNA viruses, bunyaviruses possess a nucleocapsid protein (NP) to facilitate genomic RNA encapsidation and virus replication. The structures of two NPs of members of different genera within the Bunyaviridae family have been reported. However, their structures, RNA-binding features, and functions beyond RNA binding significantly differ from one another. Here, we report the crystal structure of the BUNV NP-RNA complex. The polypeptide of the BUNV NP was found to possess a distinct fold among viral NPs. An N-terminal arm and a C-terminal tail were found to interact with neighboring NP protomers to form a tetrameric ringshaped organization. Each protomer bound a 10-nt RNA molecule, which was acquired from the expression host, in the positively charged crevice between the N and C lobes. Inhomogeneous oligomerization was observed for the recombinant BUNV NP-RNA complex, which was similar to the Rift Valley fever virus NP-RNA complex. This result suggested that the flexibility of one NP protomer with adjacent protomers underlies the BUNV ribonucleoprotein complex (RNP) formation. Electron microscopy revealed that the monomer-sized NP-RNA complex was the building block of the natural BUNV RNP. Combined with previous results indicating that mutagenesis of the interprotomer or protein-RNA interface affects BUNV replication, our structure provides a great potential for understanding the mechanism underlying negative-sense single-stranded RNA RNP formation and enables the development of antiviral therapies targeting BUNV RNP formation.assembly | packaging B unyaviruses constitute the largest segmented negative-sense single-stranded RNA (-ssRNA) virus family, which is subdivided into Orthobunyavirus, Hantavirus, Nairovirus, Phlebovirus, and Tospovirus genera (1). The genomes of Bunyaviridae members are featured by three segments, the large (L), middle (M), and small (S) segments (2, 3). The L segment encodes an RNA-dependent RNA polymerase (RdRp), the M segment encodes a precursor of glycoproteins (Gn and Gc), and the S segment encodes a nucleocapsid protein (NP). Moreover, a few Bunyaviridae members possess a nonstructural protein (NSs and/or NSm) using an ambisense coding strategy by the S and M segments (4).Similar to other -ssRNA viruses, all members of the Bunyaviridae family contain a ribonucleoprotein (RNP) complex composed of genomic RNA enwrapped by the NP (4). After entry into the cytoplasm through membrane fusion mediated by glycoproteins, the RNP is released from the virion and serves as the template with which the copackaged RdRp transcribes mRNAs from the viral genome in the RNP. In the later stage of virus replication, complementary positive-strand RNA (cRNA) is produced in the form of an RNP. The RNP serves as the template for replication that generates the viral genomic RNA in the form of an RNP ready to be packaged in the virion. Throughout...

show abstract

Section: Resultssupporting

confidence: 82%

supporting

confidence: 79%

Bunyamwera virus possesses a distinct nucleocapsid protein to facilitate genome encapsidation

Li¹,

Wang

Pan

et al. 2013

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

View full text Add to dashboard Cite

show abstract

“…For most of the NSVs the RNA synthesis catalytic activity is included in the very large L protein, a multienzymatic polypeptide responsible for mRNA synthesis and modification, as well as the generation of progeny RNPs (Kranzusch and Whelan, 2012;Morin et al, 2013). As an exception, the Orthomyxoviruses have the required enzymatic activities distributed among three different polypeptides (the PB1, PB2 and PA proteins) that are tightly associated in a heterotrimer (Fodor, 2013;Martin-Benito and Ortin, 2013;Resa-Infante et al, 2011;Ruigrok et al, 2010).…”

Section: The Viral Polymerasementioning

confidence: 96%

“…Due to space limitations, many important contributions could not be directly cited. More detailed information is available in recent specific reviews (Albertini et al, 2011;Boivin et al, 2010;Eisfeld et al, 2014;Fodor, 2013;Ivanov et al, 2011;Kranzusch and Whelan, 2012;MartinBenito and Ortin, 2013;Morin et al, 2013;Reguera et al, 2014;Resa-Infante et al, 2011;Ruigrok et al, 2010;Ruigrok et al, 2011;.…”

Section: Introductionmentioning

confidence: 96%

The RNA synthesis machinery of negative-stranded RNA viruses

Ortı́n

Martín‐Benito

2015

Virology

View full text Add to dashboard Cite

The group of Negative-Stranded RNA Viruses (NSVs) includes many human pathogens, like the influenza, measles, mumps, respiratory syncytial or Ebola viruses, which produce frequent epidemics of disease and occasional, high mortality outbreaks by transmission from animal reservoirs. The genome of NSVs consists of one to several single-stranded, negative-polarity RNA molecules that are always assembled into mega Dalton-sized complexes by association to many nucleoprotein monomers. These RNA-protein complexes or ribonucleoproteins function as templates for transcription and replication by action of the viral RNA polymerase and accessory proteins. Here we review our knowledge on these large RNA-synthesis machines, including the structure of their components, the interactions among them and their enzymatic activities, and we discuss models showing how they perform the virus transcription and replication programmes.

show abstract

“…To our knowledge, this is the first time that adaptive mutations have influenced this particular function of matrix proteins. The suppressive effect of matrix proteins on viral replication and transcription is a well-known phenomenon (41). Cellular proteins that likely are involved in the mediation of this suppressive effect are unknown.…”

Section: Discussionmentioning

confidence: 99%

A Single Amino Acid Change in the Marburg Virus Matrix Protein VP40 Provides a Replicative Advantage in a Species-Specific Manner

Koehler

Kolesnikova

Welzel

et al. 2016

J Virol

View full text Add to dashboard Cite

Marburg virus (MARV) induces severe hemorrhagic fever in humans and nonhuman primates but only transient nonlethal disease in rodents. However, sequential passages of MARV in rodents boosts infection leading to lethal disease. Guinea pig-adapted MARV contains one mutation in the viral matrix protein VP40 at position 184 (VP40 D184N ). The contribution of the D184N mutation to the efficacy of replication in a new host is unknown. In the present study, we demonstrated that recombinant MARV containing the D184N mutation in VP40 [rMARV VP40(D184N) ] grew to higher titers than wild-type recombinant MARV (rMARV WT ) in guinea pig cells. Moreover, rMARV VP40(D184N) displayed higher infectivity in guinea pig cells. Comparative analysis of VP40 functions indicated that neither the interferon (IFN)-antagonistic function nor the membrane binding capabilities of VP40 were affected by the D184N mutation. However, the production of VP40-induced virus-like particles (VLPs) and the recruitment of other viral proteins to the budding site was improved by the D184N mutation in guinea pig cells, which resulted in the higher infectivity of VP40 D184N -induced infectious VLPs (iVLPs) compared to that of VP40-induced iVLPs. In addition, the function of VP40 in suppressing viral RNA synthesis was influenced by the D184N mutation specifically in guinea pig cells, thus allowing greater rates of transcription and replication. Our results showed that the improved viral fitness of rMARV VP40(D184N) in guinea pig cells was due to the better viral assembly function of VP40 D184N and its lower inhibitory effect on viral transcription and replication rather than modulation of the VP40-mediated suppression of IFN signaling. IMPORTANCEThe increased virulence achieved by virus passaging in a new host was accompanied by mutations in the viral genome. Analyzing how these mutations affect the functions of viral proteins and the ability of the virus to grow within new host cells helps in the understanding of the molecular mechanisms increasing virulence. Using a reverse genetics approach, we demonstrated that a single mutation in MARV VP40 detected in a guinea pig-adapted MARV provided a replicative advantage of rMARV VP40(D184N) in guinea pig cells. Our studies show that this replicative advantage of rMARV VP40 D184N was based on the improved functions of VP40 in iVLP assembly and in the regulation of transcription and replication rather than on the ability of VP40 to combat the host innate immunity. F iloviruses, including Ebolaviruses (EBOV) and Marburg virus (MARV), are enveloped, nonsegmented, negative-strand RNA viruses (1). These viruses are known to cause severe fevers in humans and nonhuman primates, with case fatality rates of up to 90% (2). Although several antivirals and vaccines currently are being tested in clinical studies, none of them are licensed for human use. Therefore, work with filoviruses is restricted to biosafety level 4 (BSL-4) facilities. The recent EBOV outbreak in Guinea, Sierra Leone, and Liberia demonstrated the ...

show abstract

Architecture and regulation of negative-strand viral enzymatic machinery

Cited by 29 publications

References 106 publications

Bunyamwera virus possesses a distinct nucleocapsid protein to facilitate genome encapsidation

Bunyamwera virus possesses a distinct nucleocapsid protein to facilitate genome encapsidation

The RNA synthesis machinery of negative-stranded RNA viruses

A Single Amino Acid Change in the Marburg Virus Matrix Protein VP40 Provides a Replicative Advantage in a Species-Specific Manner

Contact Info

Product

Resources

About