Structure of Severe Fever with Thrombocytopenia Syndrome Virus Nucleocapsid Protein in Complex with Suramin Reveals Therapeutic Potential

Jiao, Libin; Ouyang, Songying; Liang, Mifang; Niu, Fujun; Shaw, Neil; Wu, Wei; Ding, Wei; Jin, Cong; Peng, Yao; Zhu, Yanping; Zhang, Fu‐Shun; Wang, Tao; Li, Chuan; Zuo, Xiaobing; Luan, Chi Hao; Li, Dexin; Liu, Zhi Jie

doi:10.1128/jvi.00672-13

Cited by 67 publications

(49 citation statements)

References 44 publications

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“…The atomic structures of nucleocapsid-like particles (NLPs) have been reported for three NSV families: Rhabdoviridae, Paramyxoviridae (genus Pneumovirus), and Bunyaviridae (genera Phlebovirus and Orthobunyavirus) (11)(12)(13)(14)(27)(28)(29)(30)(31). A comparison of representative structures from each genus was performed.…”

Section: Resultsmentioning

confidence: 99%

Common Mechanism for RNA Encapsidation by Negative-Strand RNA Viruses

Green

Cox

Tsao

et al. 2014

J Virol

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The nucleocapsid of a negative-strand RNA virus is assembled with a single nucleocapsid protein and the viral genomic RNA. The nucleocapsid protein polymerizes along the length of the single-strand genomic RNA (viral RNA) or its cRNA. This process of encapsidation occurs concomitantly with genomic replication. Structural comparisons of several nucleocapsid-like particles show that the mechanism of RNA encapsidation in negative-strand RNA viruses has many common features. Fundamentally, there is a unifying mechanism to keep the capsid protein protomer monomeric prior to encapsidation of viral RNA. In the nucleocapsid, there is a cavity between two globular domains of the nucleocapsid protein where the viral RNA is sequestered. The viral RNA must be transiently released from the nucleocapsid in order to reveal the template RNA sequence for transcription/ replication. There are cross-molecular interactions among the protein subunits linearly along the nucleocapsid to stabilize its structure. Empty capsids can form in the absence of RNA. The common characteristics of RNA encapsidation not only delineate the evolutionary relationship of negative-strand RNA viruses but also provide insights into their mechanism of replication. IMPORTANCEWhat separates negative-strand RNA viruses (NSVs) from the rest of the virosphere is that the nucleocapsid of NSVs serves as the template for viral RNA synthesis. Their viral RNA-dependent RNA polymerase can induce local conformational changes in the nucleocapsid to temporarily release the RNA genome so that the viral RNA-dependent RNA polymerase can use it as the template for RNA synthesis during both transcription and replication. After RNA synthesis at the local region is completed, the viral RNA-dependent RNA polymerase processes downstream, and the RNA genome is restored in the nucleocapsid. We found that the nucleocapsid assembly of all NSVs shares three essential elements: a monomeric capsid protein protomer, parallel orientation of subunits in the linear nucleocapsid, and a (5H ؉ 3H) motif that forms a proper cavity for sequestration of the RNA. This observation also suggests that all NSVs evolved from a common ancestor that has this unique nucleocapsid.

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Section: Resultsmentioning

confidence: 99%

Common Mechanism for RNA Encapsidation by Negative-Strand RNA Viruses

Green

Cox

Tsao

et al. 2014

J Virol

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“…Human cystatin C and the activated cathepsins were mixed in the assay buffer for 1 min, 5 min, 30 min, 1 h and 4 h at 16 °C or 25 °C and analyzed using SDS-PAGE analysis. npg NaCl) were collected on the SIBYLS beamline at the Advanced Light Source (ALS), Lawrence Berkeley National Laboratory, and the data were treated as previously described [44,45]. Briefly, each sample was measured at three exposures (0.5, 1.0, and 6.0 sec) and three concentrations (2.5, 5.0, and 10.0 mg/ml) at 10 °C to exclude aggregation and radiation-damage effects.…”

Section: Autoactivation Of Aep and Enzymatic Activity Assaymentioning

confidence: 99%

Structural analysis of asparaginyl endopeptidase reveals the activation mechanism and a reversible intermediate maturation stage

et al. 2014

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Asparaginyl endopeptidase (AEP) is an endo/lysosomal cysteine endopeptidase with a preference for an asparagine residue at the P1 site and plays an important role in the maturation of toll-like receptors 3/7/9. AEP is known to undergo autoproteolytic maturation at acidic pH for catalytic activation. Here, we describe crystal structures of the AEP proenzyme and the mature forms of AEP. Structural comparisons between AEP and caspases revealed similarities in the composition of key residues and in the catalytic mechanism. Mutagenesis studies identified N44, R46, H150, E189, C191, S217/S218 and D233 as residues that are essential for the cleavage of the peptide substrate. During maturation, autoproteolytic cleavage of AEP's cap domain opens up access to the active site on the core domain. Unexpectedly, an intermediate autoproteolytic maturation stage was discovered at approximately pH 4.5 in which the partially activated AEP could be reversed back to its proenzyme form. This unique feature was confirmed by the crystal structure of AEP pH4.5 (AEP was matured at pH 4.5 and crystallized at pH 8.5), in which the broken peptide bonds were religated and the structure was transformed back to its proenzyme form. Additionally, the AEP inhibitor cystatin C could be digested by the fully activated AEP, but could not be digested by activated cathepsins. Thus, we demonstrate for the first time that cystatins may regulate the activity of AEP through substrate competition for the active site.

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“…Since 2010, a series of structural studies, including studies on the structures of NPs encoded by phleboviruses (Rift Valley fever virus [RVFV] [11][12][13] and severe fever with thrombocytopenia syndrome virus [SFTSV] [14]), nairovirus (CrimeanCongo hemorrhagic fever virus [CCHFV]) (6,15,16), and orthobunyaviruses (Bunyamwera virus [BUNV], etc.) (17)(18)(19)(20)(21), has advanced the understanding of the structures and functions of bunyavirus NPs.…”

mentioning

confidence: 99%

Crystal Structure of the Core Region of Hantavirus Nucleocapsid Protein Reveals the Mechanism for Ribonucleoprotein Complex Formation

Guo

Wang

Sun

et al. 2016

J Virol

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Hantaviruses, which belong to the genus Hantavirus in the family Bunyaviridae, infect mammals, including humans, causing either hemorrhagic fever with renal syndrome (HFRS) or hantavirus cardiopulmonary syndrome (HCPS) in humans with high mortality. Hantavirus encodes a nucleocapsid protein (NP) to encapsidate the genome and form a ribonucleoprotein complex (RNP) together with viral polymerase. Here, we report the crystal structure of the core domains of NP (NP core ) encoded by Sin Nombre virus (SNV) and Andes virus (ANDV), which are two representative members that cause HCPS in the New World. The constructs of SNV and ANDV NP core exclude the N-and C-terminal portions of full polypeptide to obtain stable proteins for crystallographic study. The structure features an N lobe and a C lobe to clamp RNA-binding crevice and exhibits two protruding extensions in both lobes. The positively charged residues located in the RNA-binding crevice play a key role in RNA binding and virus replication. We further demonstrated that the C-terminal helix and the linker region connecting the N-terminal coiled-coil domain and NP core are essential for hantavirus NP oligomerization through contacts made with two adjacent protomers. Moreover, electron microscopy (EM) visualization of native RNPs extracted from the virions revealed that a monomer-sized NP-RNA complex is the building block of viral RNP. This work provides insight into the formation of hantavirus RNP and provides an understanding of the evolutionary connections that exist among bunyaviruses. IMPORTANCEHantaviruses are distributed across a wide and increasing range of host reservoirs throughout the world. In particular, hantaviruses can be transmitted via aerosols of rodent excreta to humans or from human to human and cause HFRS and HCPS, with mortalities of 15% and 50%, respectively. Hantavirus is therefore listed as a category C pathogen. Hantavirus encodes an NP that plays essential roles both in RNP formation and in multiple biological functions. NP is also the exclusive target for the serological diagnoses. This work reveals the structure of hantavirus NP, furthering the knowledge of hantavirus RNP formation, revealing the relationship between hantavirus NP and serological specificity and raising the potential for the development of new diagnosis and therapeutics targeting hantavirus infection. Hantavirus causes severe infectious diseases in human and animals (1, 2). In addition to 24 representative species, increasing numbers of antigenically and genetically distinct hantaviruses have recently been registered as new members of the genus Hantavirus (3, 4). The reservoir hosts of hantaviruses in nature are primarily rodents, shrews, moles, and bats, and the transmission of hantaviruses via aerosols of rodent excreta to humans causes two human diseases: (i) hemorrhagic fever with renal syndrome (HFRS), which is primarily caused by Hantaan virus (HTNV) in Asia, by Puumala virus (PUUV) and Dobrava virus (DOBV) in Europe, and by Seoul virus (SEOV) worldwide, and ...

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Structure of Severe Fever with Thrombocytopenia Syndrome Virus Nucleocapsid Protein in Complex with Suramin Reveals Therapeutic Potential

Cited by 67 publications

References 44 publications

Common Mechanism for RNA Encapsidation by Negative-Strand RNA Viruses

Common Mechanism for RNA Encapsidation by Negative-Strand RNA Viruses

Structural analysis of asparaginyl endopeptidase reveals the activation mechanism and a reversible intermediate maturation stage

Crystal Structure of the Core Region of Hantavirus Nucleocapsid Protein Reveals the Mechanism for Ribonucleoprotein Complex Formation

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