About: Human Coronavirus NL63 Open Reading Frame 3 encodes a virion-incorporated N-glycosylated membrane protein

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About: Human Coronavirus NL63 Open Reading Frame 3 encodes a virion-incorporated N-glycosylated membrane protein Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : covidontheweb.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Human Coronavirus NL63 Open Reading Frame 3 encodes a virion-incorporated N-glycosylated membrane protein
Creator	Drosten, Christian Kallies, Stephan Suliman, Tasnim Niedrig, Matthias Bader, Oliver Lehmann, Dörte Voss, Daniel Fielding, Burtram Müller, Marcel Schulz, Axel Van Der Hoek, Lia
topic	covid:aa8a9f4a432fa52294b1fa674b3ffbeadeff563f#this
Source	Medline; PMC
abstract	BACKGROUND: Human pathogenic coronavirus NL63 (hCoV-NL63) is a group 1 (alpha) coronavirus commonly associated with respiratory tract infections. In addition to known non-structural and structural proteins all coronaviruses have one or more accessory proteins whose functions are mostly unknown. Our study focuses on hCoV-NL63 open reading frame 3 (ORF 3) which is a highly conserved accessory protein among coronaviruses. RESULTS: In-silico analysis of the 225 amino acid sequence of hCoV-NL63 ORF 3 predicted a triple membrane-spanning protein. Expression in infected CaCo-2 and LLC-MK2 cells was confirmed by immunofluorescence and Western blot analysis. The protein was detected within the endoplasmatic reticulum/Golgi intermediate compartment (ERGIC) where coronavirus assembly and budding takes place. Subcellular localization studies using recombinant ORF 3 protein transfected in Huh-7 cells revealed occurrence in ERGIC, Golgi- and lysosomal compartments. By fluorescence microscopy of differently tagged envelope (E), membrane (M) and nucleocapsid (N) proteins it was shown that ORF 3 protein colocalizes extensively with E and M within the ERGIC. Using N-terminally FLAG-tagged ORF 3 protein and an antiserum specific to the C-terminus we verified the proposed topology of an extracellular N-terminus and a cytosolic C-terminus. By in-vitro translation analysis and subsequent endoglycosidase H digestion we showed that ORF 3 protein is N-glycosylated at the N-terminus. Analysis of purified viral particles revealed that ORF 3 protein is incorporated into virions and is therefore an additional structural protein. CONCLUSIONS: This study is the first extensive expression analysis of a group 1 hCoV-ORF 3 protein. We give evidence that ORF 3 protein is a structural N-glycosylated and virion-incorporated protein.
has issue date	2010-01-15(xsd:dateTime)
bibo:doi	10.1186/1743-422x-7-6
bibo:pmid	20078868
has license	cc-by
sha1sum (hex)	aa8a9f4a432fa52294b1fa674b3ffbeadeff563f
schema:url	https://doi.org/10.1186/1743-422x-7-6
resource representing a document's title	Human Coronavirus NL63 Open Reading Frame 3 encodes a virion-incorporated N-glycosylated membrane protein
has PubMed Central identifier	PMC2819038
has PubMed identifier	20078868
schema:publication	Virol J
resource representing a document's body	covid:aa8a9f4a432fa52294b1fa674b3ffbeadeff563f#body_text
is http://vocab.deri.ie/void#inDataset of	https://covidontheweb.inria.fr:4443/about/id/http/ns.inria.fr/covid19/aa8a9f4a432fa52294b1fa674b3ffbeadeff563f
is schema:about of	named entity 'virions' named entity 'antiserum' named entity 'ORF' named entity 'verified' named entity 'encodes' named entity 'PROTEINS' named entity 'DETECTED' named entity 'FLAG-TAGGED' named entity 'KNOWN' named entity 'HUMAN' named entity 'COMPARTMENTS' named entity 'RESULTS' named entity 'MEMBRANE' named entity 'CONSERVED' named entity 'VIRAL PARTICLES' named entity 'PURIFIED' named entity 'SUBCELLULAR LOCALIZATION' named entity 'COMPARTMENT' named entity 'TRIPLE' named entity 'RECOMBINANT' named entity 'ACCESSORY' named entity 'REVEALED' named entity 'STRUCTURAL PROTEIN' named entity 'SPECIFIC' named entity 'TAGGED' named entity 'PATHOGENIC' named entity 'NL63' named entity 'ADDITIONAL' named entity 'IS A' named entity 'BUDDING' named entity 'BACKGROUND' named entity 'CELLS' named entity 'OCCURRENCE' named entity 'RETICULUM' named entity 'STRUCTURAL PROTEINS' named entity 'ANALYSIS' named entity 'UNKNOWN' named entity 'C-TERMINUS' named entity '225' named entity 'HUH-7' named entity 'HAVE' named entity 'GROUP 1' named entity 'STUDIES' named entity 'ANTISERUM' named entity 'PLACE' named entity 'N-TERMINUS' named entity 'IN-VITRO' named entity 'CYTOSOLIC' named entity 'INTERMEDIATE' named entity 'TRANSLATION' named entity 'ENDOGLYCOSIDASE' named entity 'GLYCOSYLATED' named entity 'CORONAVIRUS' named entity 'GOLGI' named entity 'AMINO ACID SEQUENCE' named entity 'ASSOCIATED WITH' named entity 'STRUCTURAL' named entity 'VIRIONS' named entity 'MEMBRANE PROTEIN' named entity 'VIRION' named entity 'ENVELOPE' named entity 'LYSOSOMAL' named entity 'GLYCOSYLATED' named entity 'OPEN READING FRAME' named entity 'ASSEMBLY' named entity 'PREDICTED' named entity 'TOPOLOGY' named entity 'USING' named entity 'ORF'

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