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

Détails

ID Serval
serval:BIB_19C74056FD11
Type
Actes de conférence (partie): contribution originale à la littérature scientifique, publiée à l'occasion de conférences scientifiques, dans un ouvrage de compte-rendu (proceedings), ou dans l'édition spéciale d'un journal reconnu (conference proceedings).
Collection
Publications
Titre
Molecular determinants for membrane association of the hepatitis C virus NS2 protease domain
Titre de la conférence
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
Auteur(s)
Lange C.M., Bellecave P., Kennel A., Penin F., Moradpour D., Gouttenoire J.
Adresse
Interlaken, Switzerland, September 20-21, 2012
ISBN
1424-7860
ISSN-L
0036-7672
Statut éditorial
Publié
Date de publication
2012
Volume
142
Série
Swiss Medical Weekly
Pages
3S
Langue
anglais
Résumé
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.
Création de la notice
14/02/2013 14:42
Dernière modification de la notice
20/08/2019 13:50
Données d'usage