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  1. Article: Functional assessment of cell entry and receptor use for merbecoviruses.

    Letko, Michael

    bioRxiv : the preprint server for biology

    2024  

    Abstract: The merbecovirus subgenus of coronaviruses includes Middle East Respiratory Syndrome Coronavirus (MERS-CoV), which is a zoonotic respiratory pathogen that transmits from dromedary camels to humans and causes severe respiratory disease. Viral discovery ... ...

    Abstract The merbecovirus subgenus of coronaviruses includes Middle East Respiratory Syndrome Coronavirus (MERS-CoV), which is a zoonotic respiratory pathogen that transmits from dromedary camels to humans and causes severe respiratory disease. Viral discovery efforts have uncovered hundreds of merbecoviruses in different species across multiple continents, but few of these viruses have been isolated or studied under laboratory conditions, leaving basic questions regarding their threat to humans unresolved. Viral entry into host cells is considered an early and critical step for transmission between hosts. In this study, a scalable approach to assessing novel merbecovirus cell entry was developed and used to measure receptor use across the entire merbecovirus subgenus. Merbecoviruses are sorted into four clades based on the receptor binding domain of the spike glycoprotein. Receptor tropism is clade-specific, with only one clade using DPP4 and multiple clades using ACE2, including the entire HKU5 cluster of bat coronaviruses.
    Language English
    Publishing date 2024-03-14
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2024.03.13.584892
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Functional assessment of cell entry and receptor use for merbecoviruses

    Letko, Michael C

    bioRxiv

    Abstract: The merbecovirus subgenus of coronaviruses includes Middle East Respiratory Syndrome Coronavirus (MERS-CoV), which is a zoonotic respiratory pathogen that transmits from dromedary camels to humans and causes severe respiratory disease. Viral discovery ... ...

    Abstract The merbecovirus subgenus of coronaviruses includes Middle East Respiratory Syndrome Coronavirus (MERS-CoV), which is a zoonotic respiratory pathogen that transmits from dromedary camels to humans and causes severe respiratory disease. Viral discovery efforts have uncovered hundreds of merbecoviruses in different species across multiple continents, but few of these viruses have been isolated or studied under laboratory conditions, leaving basic questions regarding their threat to humans unresolved. Viral entry into host cells is considered an early and critical step for transmission between hosts. In this study, a scalable approach to assessing novel merbecovirus cell entry was developed and used to measure receptor use across the entire merbecovirus subgenus. Merbecoviruses are sorted into four clades based on the receptor binding domain of the spike glycoprotein. Receptor tropism is clade-specific, with only one clade using DPP4 and multiple clades using ACE2, including the entire HKU5 cluster of bat coronaviruses.
    Keywords covid19
    Language English
    Publishing date 2024-03-14
    Publisher Cold Spring Harbor Laboratory
    Document type Article ; Online
    DOI 10.1101/2024.03.13.584892
    Database COVID19

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  3. Article: Functional assessment of cell entry and receptor usage for lineage B β-coronaviruses, including 2019-nCoV.

    Letko, Michael / Munster, Vincent

    bioRxiv : the preprint server for biology

    2020  

    Abstract: Over the past 20 years, several coronaviruses have crossed the species barrier into humans, causing outbreaks of severe, and often fatal, respiratory illness. Since SARS-CoV was first identified in animal markets, global viromics projects have discovered ...

    Abstract Over the past 20 years, several coronaviruses have crossed the species barrier into humans, causing outbreaks of severe, and often fatal, respiratory illness. Since SARS-CoV was first identified in animal markets, global viromics projects have discovered thousands of coronavirus sequences in diverse animals and geographic regions. Unfortunately, there are few tools available to functionally test these novel viruses for their ability to infect humans, which has severely hampered efforts to predict the next zoonotic viral outbreak. Here we developed an approach to rapidly screen lineage B betacoronaviruses, such as SARS-CoV and the recent 2019-nCoV, for receptor usage and their ability to infect cell types from different species. We show that host protease processing during viral entry is a significant barrier for several lineage B viruses and that bypassing this barrier allows several lineage B viruses to enter human cells through an unknown receptor. We also demonstrate how different lineage B viruses can recombine to gain entry into human cells and confirm that human ACE2 is the receptor for the recently emerging 2019-nCoV.
    Keywords covid19
    Language English
    Publishing date 2020-01-22
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2020.01.22.915660
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Studying Evolutionary Adaptation of MERS-CoV.

    Letko, Michael / Munster, Vincent

    Methods in molecular biology (Clifton, N.J.)

    2019  Volume 2099, Page(s) 3–8

    Abstract: ... replication on semi-permissive cell lines as described in Letko et al., Cell Rep 24, 1730-1737, 2018. ...

    Abstract Forced viral adaptation is a powerful technique employed to study the ways viruses may overcome various selective pressures that reduce viral replication. Here, we describe methods for in vitro serial passaging of Middle East respiratory syndrome coronavirus (MERS-CoV) to select for mutations which increase replication on semi-permissive cell lines as described in Letko et al., Cell Rep 24, 1730-1737, 2018.
    MeSH term(s) Adaptation, Physiological/genetics ; Animals ; Biological Evolution ; Cell Line ; Chlorocebus aethiops ; Coronavirus Infections/virology ; Host Specificity ; Humans ; Middle East Respiratory Syndrome Coronavirus/genetics ; Middle East Respiratory Syndrome Coronavirus/physiology ; Serial Passage ; Vero Cells ; Virus Replication/genetics
    Keywords covid19
    Language English
    Publishing date 2019-12-25
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Intramural
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-0716-0211-9_1
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: A sarbecovirus found in Russian bats uses human ACE2

    Seifert, Stephanie N. / Letko, Michael C

    bioRxiv

    Abstract: Spillover of sarbecoviruses from animals to humans resulted in outbreaks of severe acute respiratory syndrome SARS-CoVs and the ongoing COVID-19 pandemic. Efforts to identify the origins of SARS-CoV-1 and -2 has resulted in the discovery of numerous ... ...

    Abstract Spillover of sarbecoviruses from animals to humans resulted in outbreaks of severe acute respiratory syndrome SARS-CoVs and the ongoing COVID-19 pandemic. Efforts to identify the origins of SARS-CoV-1 and -2 has resulted in the discovery of numerous animal sarbecoviruses -the majority of which are only distantly related to known human pathogens and do not infect human cells. The receptor binding domain (RBD) on sarbecoviruses engages receptor molecules on the host cell and mediates cell invasion. Here, we tested the receptor tropism for RBDs from two sarbecoviruses found in Russian horseshoe bats to screen cell entry. While these two viruses are in a viral lineage distinct from SARS-CoV-1 and -2, one virus was capable of using human ACE2 to facilitate cell entry. Our findings demonstrate that sarbecoviruses circulating in wildlife outside of Asia also exhibit compatibility with human ACE2 and should be taken into consideration for future universal sarbecovirus vaccine candidates.
    Keywords covid19
    Language English
    Publishing date 2021-12-07
    Publisher Cold Spring Harbor Laboratory
    Document type Article ; Online
    DOI 10.1101/2021.12.05.471310
    Database COVID19

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  6. Article ; Online: Functional assessment of cell entry and receptor usage for lineage B β-coronaviruses, including 2019-nCoV

    Michael Letko / Vincent Munster

    Abstract: AbstractOver the past 20 years, several coronaviruses have crossed the species barrier into humans, causing outbreaks of severe, and often fatal, respiratory illness. Since SARS- CoV was first identified in animal markets, global viromics projects have ... ...

    Abstract AbstractOver the past 20 years, several coronaviruses have crossed the species barrier into humans, causing outbreaks of severe, and often fatal, respiratory illness. Since SARS- CoV was first identified in animal markets, global viromics projects have discovered thousands of coronavirus sequences in diverse animals and geographic regions. Unfortunately, there are few tools available to functionally test these novel viruses for their ability to infect humans, which has severely hampered efforts to predict the next zoonotic viral outbreak. Here we developed an approach to rapidly screen lineage B betacoronaviruses, such as SARS-CoV and the recent 2019-nCoV, for receptor usage and their ability to infect cell types from different species. We show that host protease processing during viral entry is a significant barrier for several lineage B viruses and that bypassing this barrier allows several lineage B viruses to enter human cells through an unknown receptor. We also demonstrate how different lineage B viruses can recombine to gain entry into human cells and confirm that human ACE2 is the receptor for the recently emerging 2019-nCoV.
    Keywords covid19
    Publisher biorxiv
    Document type Article ; Online
    DOI 10.1101/2020.01.22.915660
    Database COVID19

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  7. Article ; Online: Functional assessment of cell entry and receptor usage for lineage B β-coronaviruses, including 2019-nCoV

    Letko, Michael / Munster, Vincent

    bioRxiv

    Keywords covid19
    Language English
    Publishing date 2020-01-22
    Publisher Cold Spring Harbor Laboratory
    Document type Article ; Online
    DOI 10.1101/2020.01.22.915660
    Database COVID19

    Full text online

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    Inter-library loan at ZB MED

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  8. Article: Studying Evolutionary Adaptation of MERS-CoV

    Letko, Michael / Munster, Vincent

    Methods Mol Biol

    Abstract: ... replication on semi-permissive cell lines as described in Letko et al., Cell Rep 24, 1730-1737, 2018. ...

    Abstract Forced viral adaptation is a powerful technique employed to study the ways viruses may overcome various selective pressures that reduce viral replication. Here, we describe methods for in vitro serial passaging of Middle East respiratory syndrome coronavirus (MERS-CoV) to select for mutations which increase replication on semi-permissive cell lines as described in Letko et al., Cell Rep 24, 1730-1737, 2018.
    Keywords covid19
    Publisher WHO
    Document type Article
    Note WHO #Covidence: #710926
    Database COVID19

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  9. Article ; Online: Functional assessment of cell entry and receptor usage for SARS-CoV-2 and other lineage B betacoronaviruses.

    Letko, Michael / Marzi, Andrea / Munster, Vincent

    Nature microbiology

    2020  Volume 5, Issue 4, Page(s) 562–569

    Abstract: Over the past 20 years, several coronaviruses have crossed the species barrier into humans, causing outbreaks of severe, and often fatal, respiratory illness. Since SARS-CoV was first identified in animal markets, global viromics projects have discovered ...

    Abstract Over the past 20 years, several coronaviruses have crossed the species barrier into humans, causing outbreaks of severe, and often fatal, respiratory illness. Since SARS-CoV was first identified in animal markets, global viromics projects have discovered thousands of coronavirus sequences in diverse animals and geographic regions. Unfortunately, there are few tools available to functionally test these viruses for their ability to infect humans, which has severely hampered efforts to predict the next zoonotic viral outbreak. Here, we developed an approach to rapidly screen lineage B betacoronaviruses, such as SARS-CoV and the recent SARS-CoV-2, for receptor usage and their ability to infect cell types from different species. We show that host protease processing during viral entry is a significant barrier for several lineage B viruses and that bypassing this barrier allows several lineage B viruses to enter human cells through an unknown receptor. We also demonstrate how different lineage B viruses can recombine to gain entry into human cells, and confirm that human ACE2 is the receptor for the recently emerging SARS-CoV-2.
    MeSH term(s) Angiotensin-Converting Enzyme 2 ; Animals ; Betacoronavirus/chemistry ; Betacoronavirus/classification ; Betacoronavirus/physiology ; CD13 Antigens/metabolism ; COVID-19 ; Cell Line ; Coronavirus Infections/metabolism ; Coronavirus Infections/virology ; Dipeptidyl Peptidase 4/metabolism ; Humans ; Mutation ; Pandemics ; Peptidyl-Dipeptidase A/chemistry ; Peptidyl-Dipeptidase A/genetics ; Peptidyl-Dipeptidase A/metabolism ; Pneumonia, Viral/metabolism ; Pneumonia, Viral/virology ; Protein Domains ; Receptors, Coronavirus ; Receptors, Virus/chemistry ; Receptors, Virus/genetics ; Receptors, Virus/metabolism ; Recombinant Fusion Proteins/metabolism ; SARS Virus/chemistry ; SARS Virus/physiology ; SARS-CoV-2 ; Spike Glycoprotein, Coronavirus/chemistry ; Spike Glycoprotein, Coronavirus/genetics ; Spike Glycoprotein, Coronavirus/metabolism ; Trypsin/metabolism ; Virus Internalization
    Chemical Substances Receptors, Coronavirus ; Receptors, Virus ; Recombinant Fusion Proteins ; Spike Glycoprotein, Coronavirus ; spike glycoprotein, SARS-CoV ; spike protein, SARS-CoV-2 ; CD13 Antigens (EC 3.4.11.2) ; DPP4 protein, human (EC 3.4.14.5) ; Dipeptidyl Peptidase 4 (EC 3.4.14.5) ; Peptidyl-Dipeptidase A (EC 3.4.15.1) ; ACE2 protein, human (EC 3.4.17.23) ; Angiotensin-Converting Enzyme 2 (EC 3.4.17.23) ; Trypsin (EC 3.4.21.4)
    Keywords covid19
    Language English
    Publishing date 2020-02-24
    Publishing country England
    Document type Journal Article ; Research Support, N.I.H., Intramural
    ISSN 2058-5276
    ISSN (online) 2058-5276
    DOI 10.1038/s41564-020-0688-y
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article ; Online: Sequence determinants of human-cell entry identified in ACE2-independent bat sarbecoviruses: A combined laboratory and computational network science approach.

    Khaledian, Ehdieh / Ulusan, Sinem / Erickson, Jeffery / Fawcett, Stephen / Letko, Michael C / Broschat, Shira L

    EBioMedicine

    2022  Volume 79, Page(s) 103990

    Abstract: Background: The sarbecovirus subgenus of betacoronaviruses is widely distributed throughout bats and other mammals globally and includes human pathogens, SARS-CoV and SARS-CoV-2. The most studied sarbecoviruses use the host protein, ACE2, to infect ... ...

    Abstract Background: The sarbecovirus subgenus of betacoronaviruses is widely distributed throughout bats and other mammals globally and includes human pathogens, SARS-CoV and SARS-CoV-2. The most studied sarbecoviruses use the host protein, ACE2, to infect cells. Curiously, the majority of sarbecoviruses identified to date do not use ACE2 and cannot readily acquire ACE2 binding through point mutations. We previously screened a broad panel of sarbecovirus spikes for cell entry and observed bat-derived viruses that could infect human cells, independent of ACE2. Here we further investigate the sequence determinants of cell entry for ACE2-independent bat sarbecoviruses.
    Methods: We employed a network science-based approach to visualize sequence and entry phenotype similarities across the diversity of sarbecovirus spike protein sequences. We then verified these computational results and mapped determinants of viral entry into human cells using recombinant chimeric spike proteins within an established viral pseudotype assay.
    Findings: We show ACE2-independent viruses that can infect human and bat cells in culture have a similar putative receptor binding motif, which can impart human cell entry into other bat sarbecovirus spikes that cannot otherwise infect human cells. These sequence determinants of human cell entry map to a surface-exposed protrusion from the predicted bat sarbecovirus spike receptor binding domain structure.
    Interpretation: Our findings provide further evidence of a group of bat-derived sarbecoviruses with zoonotic potential and demonstrate the utility in applying network science to phenotypic mapping and prediction.
    Funding: This work was supported by Washington State University and the Paul G. Allen School for Global Health.
    MeSH term(s) Angiotensin-Converting Enzyme 2/genetics ; Animals ; COVID-19 ; Chiroptera ; Humans ; Severe acute respiratory syndrome-related coronavirus ; SARS-CoV-2 ; Spike Glycoprotein, Coronavirus/metabolism ; Virus Internalization
    Chemical Substances Spike Glycoprotein, Coronavirus ; spike protein, SARS-CoV-2 ; Angiotensin-Converting Enzyme 2 (EC 3.4.17.23)
    Language English
    Publishing date 2022-04-08
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 2851331-9
    ISSN 2352-3964
    ISSN (online) 2352-3964
    DOI 10.1016/j.ebiom.2022.103990
    Database MEDical Literature Analysis and Retrieval System OnLINE

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