Molecular determinants for membrane association of the hepatitis C virus NS2 protease domain

Details

Serval ID
serval:BIB_19C74056FD11
Type
Inproceedings: an article in a conference proceedings.
Collection
Publications
Title
Molecular determinants for membrane association of the hepatitis C virus NS2 protease domain
Title of the conference
Annual Meeting of the Swiss Society of Gastroenterology, Swiss Society of Visceral Surgery, Swiss Association of the Study of the Liver and Swiss Society of Clinical Nutrition
Author(s)
Lange C.M., Bellecave P., Kennel A., Penin F., Moradpour D., Gouttenoire J.
Address
Interlaken, Switzerland, September 20-21, 2012
ISBN
1424-7860
ISSN-L
0036-7672
Publication state
Published
Issued date
2012
Volume
142
Series
Swiss Medical Weekly
Pages
3S
Language
english
Abstract
Background: Hepatitis C virus (HCV) nonstructural protein 2 (NS2)
plays essential roles in particle assembly and polyprotein processing.
It harbors an N-terminal membrane domain comprising three putative
transmembrane s egments ( amino acids [aa] 1-93) a nd a C-terminal
cysteine protease domain (aa 94-217). Given that the latter has been
predicted to be membrane-associated, we aimed to identify molecular
determinants for membrane association of the NS2 protease domain.
Methods: A comprehensive panel of NS2 deletion constructs was
analyzed by fluorescence microscopy, selective membrane extraction,
and m embrane flotation assays. Candidate aa r esidues involved in
membrane association were substituted by site-directed mutagenesis.
Results: The NS2 protease domain alone was found to associate with
membranes. Two N-terminal α-helices comprising aa 102-114 and aa
123-136 were found to m ediate this a ssociation, w ith c onserved
hydrophobic and positively charged aa residues representing the key
determinants. I nterestingly, m utagenesis analyses r evealed that
electrostatic interactions involving a positively charged aa residue in
α-helix aa 123-136 are required for membrane association. Mono- and
bicistronic (i.e. NS2 c leavage-independent) HCV constructs were
prepared to i nvestigate the effect o f these substitutions on RNA
replication and infectious viral particle formation.
Conclusions: T he NS2 protease d omain itself harbors m olecular
determinants for membrane association within α-helices aa 102-114
and aa 1 23-136 which may contribute to p roper p ositioning of t he
active site. These results provide new insights i nto the membrane
topology and t he p oorly understood f unction of t his essential viral
protease.
Create date
14/02/2013 13:42
Last modification date
20/08/2019 12:50
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