Inhibition of Chikungunya virus infection in cultured human muscle cells by furin inhibitors: impairment of the maturation of the E2 surface glycoprotein.
Chikungunya virus (CHIKV) is a mosquito-transmitted Alphavirus that causes in humans an acute infection characterized by polyarthralgia, fever, myalgia, and headache. Since 2005 this virus has been responsible for an epidemic outbreak of unprecedented magnitude. By analogy with other alphaviruses, it is thought that cellular proteases are able to process the viral precursor protein E3E2 to produce the receptor-binding E2 protein that associates as a heterodimer with E1. Destabilization of the heterodimer by exposure to low pH allows viral fusion and infection. We show that among a large panel of proprotein convertases, membranous furin but also PC5B can process E3E2 from African CHIKV strains at the HRQRR(64) / ST site, whereas a CHIKV strain of Asian origin is cleaved at RRQRR(64) / SI by membranous and soluble furin, PC5A, PC5B, and PACE4 but not by PC7 or SKI-1. Using fluorogenic model peptides and recombinant convertases, we observed that the Asian strain E3E2 model peptide is cleaved most efficiently by furin and PC5A. This cleavage was also observed in CHIKV-infected cells and could be blocked by furin inhibitor decanoyl-RVKR-chloromethyl ketone. This inhibitor was compared with chloroquine for its ability to inhibit CHIKV spreading in myoblast cell cultures, a cell-type previously described as a natural target of this virus. Our results demonstrate the role of furin-like proteases in the processing of CHIKV particles and point out new approaches to inhibit this infection.
Ozden
Simona
S
Unité d'Epidémiologie et Physiopathologie des Virus Oncogènes, CNRS URA 3015, Institut Pasteur, 75724 Paris, France.
Lucas-Hourani
Marianne
M
Ceccaldi
Pierre-Emmanuel
PE
Basak
Ajoy
A
Valentine
Menogh
M
Benjannet
Suzanne
S
Hamelin
Josée
J
Jacob
Yves
Y
Mamchaoui
Kamel
K
Mouly
Vincent
V
Desprès
Philippe
P
Gessain
Antoine
A
Butler-Browne
Gillian
G
Chrétien
Michel
M
Tangy
Frédéric
F
Vidalain
Pierre-Olivier
PO
Seidah
Nabil G
NG
D018354:Alphavirus Infections / Q000201:enzymology* / Q000378:metabolism / Q000821:virology*; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000962:Antimalarials / Q000494:pharmacology; D002460:Cell Line; D002646:Chikungunya virus / Q000187:drug effects / Q000502:physiology*; D002738:Chloroquine / Q000494:pharmacology; D045683:Furin / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D006801:Humans; D032446:Myoblasts / Q000166:cytology / Q000821:virology*; D043484:Proprotein Convertases / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D014759:Viral Envelope Proteins / Q000378:metabolism*; D053586:Virus Internalization / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects
D000590:Amino Acid Chloromethyl Ketones; D000962:Antimalarials; D011500:Protein Synthesis Inhibitors; D014759:Viral Envelope Proteins; C460143:decanoylRVKRchloromethylketone; D002738:Chloroquine; D043484:Proprotein Convertases; D045683:Furin