About: Assessing activity of Hepatitis A virus 3C protease using a cyclized luciferase-based biosensor

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About: Assessing activity of Hepatitis A virus 3C protease using a cyclized luciferase-based biosensor 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
title	Assessing activity of Hepatitis A virus 3C protease using a cyclized luciferase-based biosensor
Creator	Fang, Liurong Wang, Dang Xiao, Shaobo Zhu, Xinyu Chen, Jiyao Yang, Yuting Ye, Xu Lv, Mengting Zhou, Junwei Luo, Chuanzhen Xi, Yongqiang
topic	covid:a27468b9db8aa0e958968db14050a550f4fb27a7#this
source	Elsevier; Medline; PMC
abstract	Abstract Hepatitis A is an acute infection caused by Hepatitis A virus (HAV), which is widely distributed throughout the world. The HAV 3C cysteine protease (3Cpro), an important nonstructural protein, is responsible for most cleavage within the viral polyprotein and is critical for the processes of viral replication. Our group has previously demonstrated that HAV 3Cpro cleaves human NF-κB essential modulator (NEMO), a kinase required in interferon signaling. Based on this finding, we generated four luciferase-based biosensors containing the NEMO sequence (PVLKAQ↓ADIYKA) that is cleaved by HAV 3Cpro and/or the Nostoc punctiforme DnaE intein, to monitor the activity of HAV 3Cpro in human embryonic kidney cells (HEK-293T). Western blotting showed that HAV 3Cpro recognized and cleaved the NEMO cleavage sequence incorporated in the four biosensors, whereas only one cyclized luciferase-based biosensor (233-DnaE-HAV, 233DH) showed a measurable and reliable increase in firefly luciferase activity, with very low background, in the presence of HAV 3Cpro. With this biosensor (233DH), we monitored HAV 3Cpro activity in HEK-293T cells, and tested it against a catalytically deficient mutant HAV 3Cpro and other virus-encoded proteases. The results showed that the activity of this luciferase biosensor is specifically dependent on HAV 3Cpro. Collectively, our data demonstrate that the luciferase biosensor developed here might provide a rapid, sensitive, and efficient evaluation of HAV 3Cpro activity, and should extend our better understanding of the biological relevance of HAV 3Cpro.
has issue date	2017-07-08(xsd:dateTime)
bibo:doi	10.1016/j.bbrc.2017.05.063
bibo:pmid	28501618
has license	els-covid
sha1sum (hex)	a27468b9db8aa0e958968db14050a550f4fb27a7
schema:url	https://doi.org/10.1016/j.bbrc.2017.05.063
resource representing a document's title	Assessing activity of Hepatitis A virus 3C protease using a cyclized luciferase-based biosensor
has PubMed Central identifier	PMC7092888
has PubMed identifier	28501618
schema:publication	Biochemical and Biophysical Research Communications
resource representing a document's body	covid:a27468b9db8aa0e958968db14050a550f4fb27a7#body_text
is http://vocab.deri.ie/void#inDataset of	proxy:http/ns.inria.fr/covid19/a27468b9db8aa0e958968db14050a550f4fb27a7
is schema:about of	named entity 'Our' named entity 'Western blotting' named entity 'HEPATITIS A VIRUS' named entity 'MODULATOR' named entity 'VIRUS' named entity 'FIREFLY LUCIFERASE ACTIVITY' named entity 'EXTEND' named entity 'PRESENCE OF' named entity 'PROCESSES' named entity '293T CELLS' named entity 'EVALUATION' named entity 'CAUSED BY' named entity 'GENERATED' named entity 'LUCIFERASE' named entity 'CRITICAL' named entity 'DNAE' named entity 'FINDING' named entity 'MONITOR' named entity 'BASED' named entity 'CONTAINING' named entity 'CLEAVAGE' named entity 'BETTER' named entity 'RAPID' named entity 'WORLD' named entity 'GROUP' named entity 'DATA' named entity 'BIOLOGICAL' named entity 'HEK' named entity 'HERE' named entity 'HUMAN' named entity 'DEPENDENT' named entity 'NF-KB' named entity 'RELIABLE' named entity 'VERY LOW' named entity 'HEPATITIS A VIRUS' named entity 'RESPONSIBLE' named entity 'UNDERSTANDING' named entity 'ACUTE INFECTION' named entity 'USING' named entity 'CLEAVES' named entity 'CELLS' named entity 'INTERFERON SIGNALING' named entity 'PROVIDE' named entity 'BACKGROUND' named entity 'CLEAVED' named entity 'ENCODED' named entity 'NOSTOC PUNCTIFORME' named entity 'MUTANT' named entity 'DISTRIBUTED' named entity 'PREVIOUSLY' covid:arg/a27468b9db8aa0e958968db14050a550f4fb27a7 named entity 'SEQUENCE' named entity 'VIRAL REPLICATION' named entity 'SENSITIVE' named entity 'NEMO' named entity 'MEASURABLE' named entity 'ACTIVITY' named entity 'BIOSENSOR' named entity 'LUCIFERASE' named entity 'BASED' named entity 'EMBRYONIC KIDNEY' named entity 'TESTED' named entity 'HEPATITIS A' named entity 'BIOSENSOR' named entity '293T' named entity 'RESULTS' named entity 'ESSENTIAL' named entity 'VIRAL' named entity 'DEMONSTRATED'

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