Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the coronavirus disease-19 pandemic. One of the key components of the coronavirus replication complex are the RNA methyltransferases (MTases), RNA-modifying enzymes crucial for RNA cap formation. Recently, the structure of the 2’-O MTase has become available; however, its biological characterization within the infected cells remains largely elusive. Here, we report a novel monoclonal antibody directed against the SARS-CoV-2 non-structural protein nsp10, a subunit of both the 2’-O RNA and N7 MTase protein complexes. Using this antibody, we investigated the subcellular localization of the SARS-CoV-2 MTases in cells infected with the SARS-CoV-2.
Highlights
SARS-CoV-2 Capping EnzymesAntibodies are usually heterooligomeric glycoproteins that represent the most important components of the humoral part of the adaptive immune system
They are successful diagnostic and therapeutic tools in medical applications including fighting the coronavirus disease19 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) [3,4]
We show that the nsp10 protein is localized mainly in vesicular structures in the perinuclear region of the infected cells, where the virus is replicated
Summary
Antibodies are usually heterooligomeric glycoproteins that represent the most important components of the humoral part of the adaptive immune system They are important for neutralization of pathogens such as viruses, bacteria, parasites or fungi by interfering with the pathogen attachment to the host cell. Antibodies present powerful research tools used in many common laboratory assays, such as immunofluorescence, immunoblotting, immunoprecipitation, enzyme-linked immunosorbent assays or fluorescence-activated cell sorting They are successful diagnostic and therapeutic tools in medical applications including fighting the coronavirus disease (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus-2. We show that the antibody recognizes the nsp subunit both in its native conformation and in its denatured form Using this novel antibody, we investigated the cellular localization of nsp during cell culture infection with the SARS-CoV-2 virus. We show that the nsp protein is localized mainly in vesicular structures in the perinuclear region of the infected cells, where the virus is replicated
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