PXD033312

PXD033312 is an original dataset announced via ProteomeXchange.

Title	Integrative structural approaches identify the dynamic interactions between NS2B C-terminus and NS3 protease from DENV4 at post cleavage stage
Description	Diseases caused by flaviviruses such as dengue virus (DENV) and West Nile Virus (WNV), are a serious threat to public health. The flavivirus single-stranded RNA genome is translated into a polyprotein which is cleaved into three structural proteins and seven non-structural proteins by the viral and cellular proteases. Non-structural (NS) protein 3 is a multifunctional protein that has N-terminal protease and C-terminal helicase domains. The NS3 protease requires co-factor NS2B for enzymatic activity and folding. The NS2B-NS3 viral protease cleaves the viral polyprotein. Due to its essential role in viral replication, NS2B-NS3 protease is an attractive target for antiviral drugs. Despite the availability of crystal structures, dynamic interactions of the N- and C-termini of NS2B co-factor have been elusive due to their flexible fold. Moreover, while presence of NS2B/NS3 cleavage inhibits the activity of unlinked ZIKV NS2B-NS3 protease, no inhibition of protease activity was observed for unlinked DENV NS2B-NS3 full length with NS2B/NS3 cleavage site. In this study, we employ integrative structural approaches combined with biochemical assays to elucidate the dynamic interactions of flexible NS2B and NS3 N- and C-termini. We identified a second cis-cleavage site prior to NS2B C terminus and captured the crystal structure of self-cleaved NS2B47NS3 protease in post cleavage state. We report that the cleaved NS2B C-terminus occupy the neighbouring NS2B-NS3 molecule identifying a different mechanism where DENV NS2B C-terminus modulates NS2B-NS3 protease activity via trans-interaction in contrast to the cis-interacting NS2B C-terminus from the unlinked ZIKV protease. The intermediate conformation adopted in the reported structure can be targeted by allosteric inhibitors. The mapping of NS2B N- and C-termini with NS3 indicates that these intermolecular interactions occur mainly on the solvent-exposed beta-barrel of the NS3 protease domain. Our integrative approach enables a comprehensive understanding of the folding and dynamic interactions of DENV NS3 protease and its cofactor NS2B.
HostingRepository	jPOST
AnnounceDate	2023-04-20
AnnouncementXML	Submission_2023-04-19_08:01:15.557.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Radoslaw Sobota
SpeciesList	scientific name: cellular organisms; NCBI TaxID: 131567;
ModificationList	S-carboxamidomethyl-L-cysteine; alpha-amino acetylated residue; L-methionine sulfoxide
Instrument	instrument

Revision	Datetime	Status	ChangeLog Entry
0	2022-04-19 22:51:38	ID requested
⏵ 1	2023-04-19 08:01:15	announced

Dataset with its publication pending

submitter keyword: NS2B-NS3 protease, DENV, dengue virus, Dengue fever, viral protease, X-ray crystallography, cross linking mass spectrometry

Wint Wint Phoo
lab head
Radoslaw Sobota
contact affiliation	IMCB A-STAR Singapore
dataset submitter

jPOST dataset URI

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