RNA Synthesis and Capping by Non-segmented Negative Strand RNA Viral Polymerases: Lessons From a Prototypic Virus

Ogino, Tomoaki; Green, Todd

doi:10.3389/fmicb.2019.01490

Cited by 68 publications

(98 citation statements)

References 250 publications

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“…The nascent RNA may exit to activate the capping and methyltransferase so that the elongation of mRNA will proceed [87]. In the presence of RNA-free N, the nascent RNA can exit the L-P complex, bypassing the capping enzyme, and be encapsidated by incoming N protein subunits that can be delivered as a N-P complex to complete elongation of genome replication [88,89].…”

Section: Viral Rna Synthesismentioning

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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Section: Viral Rna Synthesismentioning

confidence: 99%

Nucleocapsid Structure of Negative Strand RNA Virus

Luo

Terrell

Mcmanus

2020

Viruses

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“…Previous studies of the NNS viral polymerase activities by others on the genomic and antigenomic promoters using cell-based minigenome assays and in vitro assays with recombinant polymerase have provided enrich insights on NNS RNA synthesis (25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40)(41)(42).…”

Section: Downloaded Frommentioning

confidence: 99%

In Vitro Primer-Based RNA Elongation and Promoter Fine Mapping of the Respiratory Syncytial Virus

Cao

Gao

Roesler

et al. 2020

J Virol

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Respiratory syncytial virus (RSV) is a nonsegmented negative-sense (NNS) RNA virus and shares a similar RNA synthesis strategy with other members of NNS RNA viruses such as measles, rabies, and Ebola. RSV RNA synthesis is catalyzed by a multifunctional RNA dependent RNA Polymerase (RdRP), which comprises of a large (L) protein that catalyzes three distinct enzymatic functions and an essential co-enzyme phosphoprotein (P). Here, we successfully prepared highly pure, full-length wild-type and mutant RSV polymerase (L:P) complex. We demonstrated that the RSV polymerase could carry out both de novo and primer-based RNA synthesis. We defined the minimal length of the RNA template for in vitro de novo RNA synthesis using the purified RSV polymerase is 8 nucleotides (nts), shorter than previously reported. We showed the RSV polymerase catalyzes the primer-dependent RNA elongation with different lengths of primers on both short (10-nt) and long (25-nt) RNA templates. We compared the sequence specificity of different viral promoters and identified positions 3, 5, and 8 of the promoter sequence are essential to the in vitro RSV polymerase activity, consistent with the results previously mapped by the in vivo minigenome assay. Overall, these findings agree well with previous biochemical studies and extend our understanding of the promoter sequence and the mechanism of RSV RNA synthesis. IMPORTANCE As a major human pathogen, respiratory syncytial virus (RSV) affects 3.4 million children worldwide annually. However, no effective antivirals or vaccines are available. An in-depth mechanistic understanding of the RSV RNA synthesis machinery remains a high priority among the nonsegmented negative-sense (NNS) RNA viruses. There is a strong public health need for research on this virus due to major fundamental gaps in our understanding of NNS RNA virus replication. As the key enzyme executing transcription and replication of the virus, the RSV RNA dependent RNA polymerase (RdRP) is a logical target for novel antiviral drugs. Therefore, exploring the primer-dependent RNA elongation extends our mechanistic understanding of the RSV RNA synthesis. Further fine mapping of the promoter sequence paves the way to better understanding the function and structure of the RSV polymerase.

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“…This assembly protects the viral genome from RNA degradation and the host's intracellular defence systems. Only the encapsidated form of viral genome can serve as a template for the transcription of mRNA or nascent replication products by the viral RNA dependent RNA polymerase (RdRp) 4 . The newly synthesized viral RNA genome is encapsidated by nascently translated N protein and directed to the viral matrix protein, located in the inner side of the host cell membrane, for virus packaging via budding 5 , to complete the viral life cycle.…”

Section: Introductionmentioning

confidence: 99%

CryoEM structure of the Nipah virus nucleocapsid assembly

Ker

Jenkins

Greive

et al. 2020

Preprint

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Nipah virus is a highly pathogenic zoonotic RNA virus, causing fatal encephalitis in humans. Like other negative-strand RNA viruses including Ebola and measles, its genome is wrapped by the nucleocapsid (N) protein forming a helical assembly. Here we report the CryoEM structure of the Nipah nucleocapsid protein-RNA assembly, at near atomic resolution. The N protein wraps the RNA genome with a periodicity of six nucleotides per protomer, around the outer edge of the helical assembly, in common with other paramyxoviruses. This structure uncovers details of the nucleocapsid assembly, demonstrating the role of the N-terminal arm of the N protein in the formation of the helical assembly and revealing details of the sequence-independent coordination of RNA binding in the "3-bases-in, 3-bases-out" conformation. CryoEM analysis also reveals formation of clam-shaped assemblies of the N-protein, mediated by intersubunit interactions involving several N protein loop regions.

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RNA Synthesis and Capping by Non-segmented Negative Strand RNA Viral Polymerases: Lessons From a Prototypic Virus

Cited by 68 publications

References 250 publications

Nucleocapsid Structure of Negative Strand RNA Virus

Nucleocapsid Structure of Negative Strand RNA Virus

In Vitro Primer-Based RNA Elongation and Promoter Fine Mapping of the Respiratory Syncytial Virus

CryoEM structure of the Nipah virus nucleocapsid assembly

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