#Replication of #MERS and #SARS #coronaviruses in #bat cells offers insights to their ancestral origins (Emerg Microbes Infect., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Emerg Microbes Infect. 2018 Dec 10;7(1):209. doi: 10.1038/s41426-018-0208-9.

Replication of MERS and SARS coronaviruses in bat cells offers insights to their ancestral origins.

Lau SKP1,2,3,4, Fan RYY5, Luk HKH5, Zhu L5, Fung J5, Li KSM5, Wong EYM5, Ahmed SS5, Chan JFW6,5,7,8, Kok RKH6,5,7,8, Chan KH6,5,7,8, Wernery U9, Yuen KY6,5,7,8, Woo PCY10,11,12,13.

Author information: 1 State Key Laboratory of Emerging Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China. skplau@hku.hk. 2 Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China. skplau@hku.hk. 3 Carol Yu Centre for Infection, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China. skplau@hku.hk. 4 Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China. skplau@hku.hk. 5 Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China. 6 State Key Laboratory of Emerging Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China. 7 Carol Yu Centre for Infection, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China. 8 Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China. 9 Central Veterinary Research Laboratory, Dubai, United Arab Emirates. 10 State Key Laboratory of Emerging Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China. pcywoo@hku.hk. 11 Department of Microbiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China. pcywoo@hku.hk. 12 Carol Yu Centre for Infection, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China. pcywoo@hku.hk. 13 Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China. pcywoo@hku.hk.



Previous findings of Middle East Respiratory Syndrome coronavirus (MERS-CoV)-related viruses in bats, and the ability of Tylonycteris-BatCoV HKU4 spike protein to utilize MERS-CoV receptor, human dipeptidyl peptidase 4 hDPP4, suggest a bat ancestral origin of MERS-CoV. We developed 12 primary bat cell lines from seven bat species, including Tylonycteris pachypus, Pipistrellus abramus and Rhinolophus sinicus (hosts of Tylonycteris-BatCoV HKU4, Pipistrellus-BatCoV HKU5, and SARS-related-CoV respectively), and tested their susceptibilities to MERS-CoVs, SARS-CoV, and human coronavirus 229E (HCoV-229E). Five cell lines, including P. abramus and R. sinicus but not T. pachypus cells, were susceptible to human MERS-CoV EMC/2012. However, three tested camel MERS-CoV strains showed different infectivities, with only two strains capable of infecting three and one cell lines respectively. SARS-CoV can only replicate in R. sinicus cells, while HCoV-229E cannot replicate in any bat cells. Bat dipeptidyl peptidase 4 (DPP4) sequences were closely related to those of human and non-human primates but distinct from dromedary DPP4 sequence. Critical residues for binding to MERS-CoV spike protein were mostly conserved in bat DPP4. DPP4 was expressed in the five bat cells susceptible to MERS-CoV, with significantly higher mRNA expression levels than those in non-susceptible cells (P = 0.0174), supporting that DPP4 expression is critical for MERS-CoV infection in bats. However, overexpression of T. pachypus DPP4 failed to confer MERS-CoV susceptibility in T. pachypus cells, suggesting other cellular factors in determining viral replication. The broad cellular tropism of MERS-CoV should prompt further exploration of host diversity of related viruses to identify its ancestral origin.

PMID: 30531999 DOI: 10.1038/s41426-018-0208-9

Keywords: Coronavirus; MERS-CoV; SARS; Bats.



Middle East respiratory syndrome #coronavirus (#MERS-CoV): #Impact on #Saudi Arabia, 2015 (Saudi J Biol Sci., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Saudi J Biol Sci. 2018 Nov;25(7):1402-1405. doi: 10.1016/j.sjbs.2016.09.020. Epub 2016 Oct 1.

Middle East respiratory syndrome coronavirus (MERS-CoV): Impact on Saudi Arabia, 2015.

Faridi U1.

Author information: 1 Department of Biochemistry, Tabuk University, Tabuk, Saudi Arabia.



Middle East respiratory syndrome is the acute respiratory syndrome caused by betacoronavirus MERS-CoV. The first case of this disease was reported from Saudi Arabia in 2012. This virus is lethal and is a close relative of a severe acute respiratory syndrome (SARS), which is responsible for more than 3000 deaths in 2002-2003. According to Ministry of Health, Saudi Arabia. The number of new cases is 457 in 2015. Riyadh has the highest number of reports in comparison to the other cities. According to this report, males are more susceptible than female, especially after the age of 40. Because of the awareness and early diagnosis the incidence is falling gradually. The pre-existence of another disease like cancer or diabetic etc. boosts the infection. MERS is a zoonotic disease and human to human transmission is low. The MERS-CoV is a RNA virus with protein envelope. On the outer surface, virus has spike like glycoprotein which is responsible for the attachment and entrance inside host cells. There is no specific treatment for the MERS-CoV till now, but drugs are in pipeline which bind with the spike glycoprotein and inhibit its entrance host cells. MERS-CoV and SAR-CoV are from the same genus, so it was thought that the drugs which inhibit the growth of SARS-CoV can also inhibit the growth of MERS-CoV but those drugs are not completely inhibiting virus activity. Until we don’t have proper structure and the treatment of MERS-CoV, We should take precautions, especially the health care workers, Camel owners and Pilgrims during Hajj and Umrah, because they are at a higher risk of getting infected.

KEYWORDS: Betacoronavirus; MERS-CoV; SARS; Saudi Arabia

PMID: 30505188 PMCID: PMC6252006  DOI: 10.1016/j.sjbs.2016.09.020

Keywords: Coronavirus; Betacoronavirus; MERS-CoV; SARS; Saudi Arabia; Human; Camels.


Molecular #identification of #Betacoronavirus in #bats from #Sardinia (#Italy): first detection and phylogeny (Virus Genes., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Virus Genes. 2018 Nov 13. doi: 10.1007/s11262-018-1614-8. [Epub ahead of print]

Molecular identification of Betacoronavirus in bats from Sardinia (Italy): first detection and phylogeny.

Lecis R1,2, Mucedda M3, Pidinchedda E3, Pittau M4,5, Alberti A4,5.

Author information: 1 Department of Veterinary Medicine, University of Sassari, Via Vienna 2, 07100, Sassari, Italy. rlecis@uniss.it. 2 Mediterranean Centre for Disease Control, University of Sassari, Via Vienna 2, 07100, Sassari, Italy. rlecis@uniss.it. 3 Centro Pipistrelli Sardegna, Via G. Leopardi 1, 07100, Sassari, Italy. 4 Department of Veterinary Medicine, University of Sassari, Via Vienna 2, 07100, Sassari, Italy. 5 Mediterranean Centre for Disease Control, University of Sassari, Via Vienna 2, 07100, Sassari, Italy.



Bats may be natural reservoirs for a large variety of emerging viruses, including mammalian coronaviruses (CoV). The recent emergence of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV) in humans, with evidence that these viruses may have their ancestry in bats, highlights the importance of virus surveillance in bat populations. Here, we report the identification and molecular characterization of a bat β-Coronavirus, detected during a viral survey carried out on different bat species in the island of Sardinia (Italy). Cutaneous, oral swabs, and faecal samples were collected from 46 bats, belonging to 15 different species, and tested for viral presence. Coronavirus RNA was detected in faecal samples from three different species: the greater horseshoe bat (Rhinolophus ferrumequinum), the brown long-eared bat (Plecotus auritus), and the European free-tailed bat (Tadarida teniotis). Phylogenetic analyses based on RNA-dependent RNA polymerase (RdRp) sequences assigned the detected CoV to clade 2b within betacoronaviruses, clustering with SARS-like bat CoVs previously reported. These findings point to the need for continued surveillance of bat CoV circulating in Sardinian bats, and extend the current knowledge on CoV ecology with novel sequences detected in bat species not previously described as β-Coronavirus hosts.

KEYWORDS: Bats; Coronavirus; RNA-dependent RNA polymerase; Rhinolophus ferrumequinum; Sardinia

PMID: 30426315 DOI: 10.1007/s11262-018-1614-8

Keywords: Coronavirus; Betacoronavirus; SARS; Bats; Italy.


#AIDS, #Avian #flu, #SARS, #MERS, #Ebola, #Zika… what next? (Vaccine, abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Vaccine. 2017 Aug 16;35(35 Pt A):4470-4474. doi: 10.1016/j.vaccine.2017.04.082. Epub 2017 Jun 19.

AIDS, Avian flu, SARS, MERS, Ebola, Zika… what next?

Reperant LA1, Osterhaus ADME2.

Author information: 1 Artemis One Health Research Foundation, Utrecht, The Netherlands. 2 Artemis One Health Research Foundation, Utrecht, The Netherlands; Research Center for Emerging Infections and Zoonoses, University of Veterinary Medicine, Hannover, Germany. Electronic address: Albert.Osterhaus@tiho-hannover.de.



Emerging infections have threatened humanity since times immemorial. The dramatic anthropogenic, behavioral and social changes that have affected humanity and the environment in the past century have accelerated the intrusion of novel pathogens into the global human population, sometimes with devastating consequences. The AIDS and influenza pandemics have claimed and will continue to claim millions of lives. The recent SARS and Ebola epidemics have threatened populations across borders. The emergence of MERS may well be warning signals of a nascent pandemic threat, while the potential for geographical spread of vector-borne diseases, such as Zika, but also Dengue and Chikungunya is unprecedented. Novel technologies and innovative approaches have multiplied to address and improve response preparedness towards the increasing yet unpredictable threat posed by emerging pathogens.

KEYWORDS: Emerging; Epidemics; Preparedness; Virus

PMID: 28633891 DOI: 10.1016/j.vaccine.2017.04.082 [Indexed for MEDLINE]

Keywords: Infectious Diseases; Emerging Diseases; Pandemic Preparedness; Zika Virus; Ebola; MERS-CoV; SARS; HIV/AIDS; Avian Influenza.


Combination #attenuation offers #strategy for live-attenuated #coronavirus #vaccines (J Virol., abstract)

[Source: Journal of Virology, full page: (LINK). Abstract, edited.]

Combination attenuation offers strategy for live-attenuated coronavirus vaccines

Vineet D. Menachery1,2, Lisa E. Gralinski2, Hugh D. Mitchell4, Kenneth H. Dinnon III2, Sarah R. Leist2, Boyd L. Yount Jr.2, Eileen T. McAnarney1,2, Rachel L. Graham2, Katrina M. Waters4 and Ralph S. Baric2,3⇑

Author Affiliations: 1 Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA; 2 Departments of Epidemiology and 3Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA; 4 Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington, USA



With an ongoing threat posed by circulating zoonotic strains, new strategies are required to prepare for the next emergent coronavirus (CoV). Previously, groups had targeted conserved coronavirus proteins as a strategy to generate live-attenuated vaccine strains against current and future CoVs. With this in mind, we explored whether manipulation of CoV NSP16, a conserved 2′O methyltransferase (MTase), could provide a broad attenuation platform against future emergent strains. Using the SARS-CoV mouse model, a NSP16 mutant vaccine was evaluated for protection from heterologous challenge, efficacy in the aging host, and potential for reversion to pathogenesis. Despite some success, concerns for virulence in the aged and potential for reversion makes targeting NSP16 alone an untenable approach. However, combining a 2′O MTase mutation with a previously described CoV fidelity mutant produced a vaccine strain capable of protection from heterologous virus challenge, efficacy in aged mice, and no evidence for reversion. Together, the results indicate that targeting the CoV 2′O MTase in parallel with other conserved attenuating mutations may provide a platform strategy for rapidly generating live-attenuated coronavirus vaccines.



Emergent coronaviruses remain a significant threat to global public health and rapid response vaccine platforms are needed to stem future outbreaks. However, failure of many previous CoV vaccine formulations has clearly highlighted the need to test efficacy under different conditions and especially in vulnerable populations like the aged and immune-compromised. This study illustrates that despite success in young models, the 2′O methyltransferase mutant carries too much risk for pathogenesis and reversion in vulnerable models to be used as a stand-alone vaccine strategy. Importantly, the 2′O methyltransferase mutation can be paired with other attenuating approaches to provide robust protection from heterologous challenge and in vulnerable populations. Coupled with increased safety and reduced pathogenesis, the study highlights the potential for 2′O methyltransferase attenuation as a major component of future live-attenuated coronavirus vaccines.



Corresponding Author: Ralph S. Baric, Address: University of North Carolina at Chapel Hill, 2107 McGavran-Greenberg Hall CB 7435, Chapel Hill, NC 27599-7435, Telephone:
919-966-7991 Fax: 919-966-0584, Email: Rbaric@email.unc.edu

Copyright © 2018 American Society for Microbiology. All Rights Reserved.

Keywords: Coronavirus; SARS; Vaccines; Animal Models.


Identification and application of self-binding zipper-like #sequences in #SARS-CoV spike protein (Int J Biochem Cell Biol., abstract)

[Source: Science Direct, full page: (LINK). Abstract, edited.]

The International Journal of Biochemistry & Cell Biology / Available online 22 May 2018 / In Press, Accepted Manuscript

Identification and application of self-binding zipper-like sequences in SARS-CoV spike protein

Si Min Zhang a, b, Ying Liao c, Tuan Ling Neo a, Yanning Lu a, Ding Xiang Liu a, Anders Vahlne b, James P. Tam a

a School of Biological Sciences, Nanyang Technological University, Singapore; b Division of Clinical Microbiology, Department of Laboratory Medicine, Karolinska Institutet, Sweden; c Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China

Received 7 December 2017, Revised 3 May 2018, Accepted 21 May 2018, Available online 22 May 2018 / DOI: https://doi.org/10.1016/j.biocel.2018.05.012



Self-binding peptides containing zipper-like sequences, such as the Leu/Ile zipper sequence within the coiled coil regions of proteins and the cross-β spine steric zippers within the amyloid-like fibrils, could bind to the protein-of-origin through homophilic sequence-specific zipper motifs. These self-binding sequences represent opportunities for the development of biochemical tools and/or therapeutics. Here, we report on the identification of a putative self-binding β-zipper-forming peptide within the severe acute respiratory syndrome-associated coronavirus spike (S) protein and its application in viral detection. Peptide array scanning of overlapping peptides covering the entire length of S protein identified 34 putative self-binding peptides of six clusters, five of which contained octapeptide core consensus sequences. The Cluster I consensus octapeptide sequence GINITNFR was predicted by the Eisenberg’s 3D profile method to have high amyloid-like fibrillation potential through steric β-zipper formation. Peptide C6 containing the Cluster I consensus sequence was shown to oligomerize and form amyloid-like fibrils. Taking advantage of this, C6 was further applied to detect the S protein expression in vitro by fluorescence staining. Meanwhile, the coiled-coil-forming Leu/Ile heptad repeat sequences within the S protein were under-represented during peptide array scanning, in agreement with that long peptide lengths were required to attain high helix-mediated interaction avidity. The data suggest that short β-zipper-like self-binding peptides within the S protein could be identified through combining the peptide scanning and predictive methods, and could be exploited as biochemical detection reagents for viral infection.

Abbreviations: S, Spike protein; HR, Heptad repeat; HIV-1, Human immunodeficiency virus type I; SARS-CoV, Severe acute respiratory syndrome associated coronavirus; RBD,  Receptor binding domain; FP, Fusion peptide; 6-HB, Six-helix bundle; MPER, Membrane proximal external region; IFP, Internal fusion peptide; HA, Hemagglutinin; PAGE, Polyacrylamide gel electrophoresis; CD, Circular dichroism; ThT, Thioflavin T

Keywords: Steric β-zipper; SARS-CoV spike protein; Class I viral fusion glycoprotein; self-binding peptides; viral detection

© 2018 Published by Elsevier Ltd.

Keywords: SARS; Coronavirus; Virology.


Broad #receptor #engagement of an emerging #global #coronavirus may potentiate its diverse cross-species #transmissibility (Proc Natl Acad Sci USA, abstract)

[Source: Proceedings of the National Academy of Sciences of the United States of America, full page: (LINK). Abstract, edited.]

Broad receptor engagement of an emerging global coronavirus may potentiate its diverse cross-species transmissibility

Wentao Li, Ruben J. G. Hulswit, Scott P. Kenney, Ivy Widjaja, Kwonil Jung, Moyasar A. Alhamo, Brenda van Dieren, Frank J. M. van Kuppeveld, Linda J. Saif, and Berend-Jan Bosch

PNAS May 14, 2018. 201802879; published ahead of print May 14, 2018. DOI: https://doi.org/10.1073/pnas.1802879115

Contributed by Linda J. Saif, April 12, 2018 (sent for review February 15, 2018; reviewed by Tom Gallagher and Stefan Pöhlmann)



Coronaviruses exhibit a propensity for interspecies transmission, with SARS- and MERS-coronaviruses as notable examples. Cross-species transmission by coronaviruses is foremost determined by the virus’ ability to bind receptors of new hosts. We here report that the recently identified, yet globally distributed porcine deltacoronavirus employs host aminopeptidase N (APN) as an entry receptor via S protein-mediated interaction with an interspecies conserved domain that allows for APN orthologue-mediated entry. Identification of APN as a deltacoronavirus receptor emphasizes the remarkable preferential employment of cell surface host peptidases as receptors by coronaviruses. Our findings provide important insight into how receptor usage of coronaviruses may fuel cross-host transmission between distantly related species and necessitate surveillance studies of deltacoronaviruses in thus far unappreciated potential reservoirs, including humans.



Porcine deltacoronavirus (PDCoV), identified in 2012, is a common enteropathogen of swine with worldwide distribution. The source and evolutionary history of this virus is, however, unknown. PDCoV belongs to the Deltacoronavirus genus that comprises predominantly avian CoV. Phylogenetic analysis suggests that PDCoV originated relatively recently from a host-switching event between birds and mammals. Insight into receptor engagement by PDCoV may shed light into such an exceptional phenomenon. Here we report that PDCoV employs host aminopeptidase N (APN) as an entry receptor and interacts with APN via domain B of its spike (S) protein. Infection of porcine cells with PDCoV was drastically reduced by APN knockout and rescued after reconstitution of APN expression. In addition, we observed that PDCoV efficiently infects cells of unusual broad species range, including human and chicken. Accordingly, PDCoV S was found to target the phylogenetically conserved catalytic domain of APN. Moreover, transient expression of porcine, feline, human, and chicken APN renders cells susceptible to PDCoV infection. Binding of PDCoV to an interspecies conserved site on APN may facilitate direct transmission of PDCoV to nonreservoir species, including humans, potentially reflecting the mechanism that enabled a virus, ancestral to PDCoV, to breach the species barrier between birds and mammals. The APN cell surface protein is also used by several members of the Alphacoronavirusgenus. Hence, our data constitute the second identification of CoVs from different genera that use the same receptor, implying that CoV receptor selection is subjected to specific restrictions that are still poorly understood.

PDCoV – spike – APN – receptor – cross-species transmission



1 W.L., R.J.G.H., and S.P.K. contributed equally to this work.

2 To whom correspondence may be addressed. Email: saif.2@osu.edu or b.j.bosch@uu.nl.

Author contributions: W.L., R.J.G.H., S.P.K., I.W., L.J.S., and B.-J.B. designed research; W.L., R.J.G.H., S.P.K., I.W., K.J., M.A.A., and B.v.D. performed research; L.J.S. contributed new reagents/analytic tools; W.L., R.J.G.H., S.P.K., I.W., K.J., M.A.A., F.J.M.v.K., L.J.S., and B.-J.B. analyzed data; and R.J.G.H. and B.-J.B. wrote the paper.

Reviewers: T.G., Loyola University Chicago; and S.P., German Primate Center.

The authors declare no conflict of interest.

This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1802879115/-/DCSupplemental.

Published under the PNAS license.

Keywords: Alphacoronavirus; Deltacoronavirus; Pigs; MERS-CoV; SARS.