Cryo-EM structure of a herpesvirus capsid at 3.1 Å

Yuan, Shuai; Wang, Jialing; Zhu, Dahuan; Wang, Nan; Gao, Qiang; Chen, Wenyuan; Tang, Hao; Wang, Junzhi; Zhang, Xinzheng; Liu, Hongrong; Rao, Zihe; Wang, Xiangxi

doi:10.1126/science.aao7283

Cited by 97 publications

(130 citation statements)

References 66 publications

(73 reference statements)

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“…1D, S1 and S2). By using the block-reconstruction method (Yuan et al, 2018;Zhu et al, 2018a), we were able to improve the resolution for the structures of the hexameric and dodecameric terminases to 3.5 Å and 3.6 Å, respectively, suggesting that the complex is intrinsically flexible with unrestrained symmetry (Figs. 1D, S2 and Table S1).…”

Section: Resultsmentioning

confidence: 99%

“…To further improve resolution we used the block-based reconstruction (Wang et al, 2018;Yuan et al, 2018;Zhu et al, 2018a;Wang et al, 2019). The orientation parameters of each particle image (hexamer and dodecamer) determined in Relion were used to guide extraction of the block region (∼50% bigger than monomer) and the block was refined and reconstructed (Scheres, 2012).…”

Section: Image Processingmentioning

confidence: 99%

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Architecture of the herpesvirus genome-packaging complex and implications for DNA translocation

et al. 2020

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Genome packaging is a fundamental process in a viral life cycle and a prime target of antiviral drugs. Herpesviruses use an ATP-driven packaging motor/terminase complex to translocate and cleave concatemeric dsDNA into procapsids but its molecular architecture and mechanism are unknown. We report atomic structures of a herpesvirus hexameric terminase complex in both the apo and ADP•BeF3-bound states. Each subunit of the hexameric ring comprises three components-the ATPase/terminase pUL15 and two regulator/fixer proteins, pUL28 and pUL33-unlike bacteriophage terminases. Distal to the nuclease domains, six ATPase domains form a central channel with conserved basicpatches conducive to DNA binding and transacting arginine fingers are essential to ATP hydrolysis and sequential DNA translocation. Rearrangement of the nuclease domains mediated by regulatory domains converts DNA translocation mode to cleavage mode. Our structures favor a sequential revolution model for DNA translocation and suggest mechanisms for concerted domain rearrangements leading to DNA cleavage. KEYWORDS dsDNA virus, genome packaging, viral maturation, terminase complex, drug target Yunxiang Yang and Pan Yang contributed equally to this work.

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Section: Resultsmentioning

confidence: 99%

Section: Image Processingmentioning

confidence: 99%

Architecture of the herpesvirus genome-packaging complex and implications for DNA translocation

et al. 2020

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“…Cryo-electron microscopy (cryo-EM) has become a robust imaging tool for determining the ultrastructure of viruses at near-atomic resolution (Yuan et al 2018). Images clearly showed four distinct electron-dense layers ( Fig.…”

Section: Virus Cultivation Purification and Morphologymentioning

confidence: 99%

Proteomic Profiling of Purified Rabies Virus Particles

et al. 2019

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While host proteins incorporated into virions during viral budding from infected cell are known to play essential roles in multiple process of the life cycle of progeny virus, these characteristics have been largely neglected in studies on rabies virus (RABV). Here, we purified the RABV virions with good purity and integrity, and analyzed their proteome by nano LC-MS/MS, followed by the confirmation with immunoblot and immuno-electronic microscopy. In addition to the 5 viral proteins, 49 cellular proteins were reproducibly identified to be incorporated into matured RABV virions. Function annotation suggested that 24 of them were likely involved in virus replication. Furthermore, cryo-EM was employed to observe the purified RABV virions, generating high-resolution pictures of the bullet-shaped virion structure of RABV. This study has provided new insights into the host proteins composition in RABV virion and shed the light for further investigation on molecular mechanisms of RABV infection, as well as the discovery of new anti-RABV therapeutics.

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“…Virion production begins in the nucleus, where capsid assembly and genome encapsidation occurs, and concludes in the cytoplasm with the envelopment of the capsid-tegument complex (5). While the structure of the capsid is solved to 3.1 Å for HSV-1 and 4.9 Å for PRV (6)(7)(8), the majority of the tegument layer departs from the radial symmetry of the capsid shell and cannot be visualized in great detail by composite averaging methods such as cryo-electron microscopy (9). This technical limitation is overcome by the application of cryo-electron tomography, but this method has yet to achieve resolutions necessary to infer protein distributions within the herpesvirus virion (10,11).…”

Section: Introductionmentioning

confidence: 99%

Dissecting the herpesvirus architecture by targeted proteolysis

Daniel

Pegg

Smith

2018

Preprint

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Herpesvirus particles have a complex architecture consisting of an icosahedral capsid that is surrounded by a lipid envelope. Connecting these two components is a layer of tegument that consists of varying amounts of twenty or more proteins. The arrangement of proteins within the tegument cannot easily be assessed and instead is inferred from tegument interactions identified in reductionist models. To better understand the tegument architecture, we have developed an approach to probe capsid-tegument interactions within extracellular viral particles by encoding tobacco etch virus (TEV) protease sites in viral structural proteins, along with distinct fluorescent tags in capsid and tegument components. In this study, TEV sites were engineered within the pUL36 large tegument protein: a critical structural element that is anchored directly on the capsid surface. Purified pseudorabies virus extracellular particles were permeabilized and TEV protease was added to selectively cleave the exposed pUL36 backbone.Interactions with the capsid were assessed in situ by monitoring the fate of the fluorescent signals following cleavage. Although several regions of pUL36 are proposed to bind capsids, pUL36 was found stably anchored to the capsid exclusively at its carboxyl terminus. Two additional tegument proteins, pUL37 and pUS3, were tethered to the capsid via pUL36 whereas the pUL16, pUL47, pUL48, and pUL49 tegument proteins were not stably bound to the capsid. IMPORTANCE:Neuroinvasive alphaherpesviruses produce diseases of clinical and economic significance in humans and veterinary animals, but are predominantly associated with less serious recurrent disease. Like all viruses, herpesviruses assemble a metastable particle that selectively dismantles during initial infection. This process is made more complex by the presence of a tegument layer that resides between the capsid surface and envelope. Components of the tegument are essential for particle assembly and also serve as critical effectors that promote infection upon entry into cells. How this dynamic network of protein interactions is arranged within virions is largely unknown. We present a molecular approach to dissect the tegument and with it, begin to tease apart the protein interactions that underlie this complex layer of the virion architecture.

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Cryo-EM structure of a herpesvirus capsid at 3.1 Å

Cited by 97 publications

References 66 publications

Architecture of the herpesvirus genome-packaging complex and implications for DNA translocation

Architecture of the herpesvirus genome-packaging complex and implications for DNA translocation

Proteomic Profiling of Purified Rabies Virus Particles

Dissecting the herpesvirus architecture by targeted proteolysis

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