Chikungunya virus requires cellular chloride channels for efficient genome replication

Abstract

Chikungunya virus (CHIKV) is a re-emerging, pathogenic alphavirus that is transmitted to humans by Aedes spp. mosquitoes—causing fever and debilitating joint pain, with frequent long-term health implications and high morbidity. The CHIKV lifecycle is poorly understood and specific antiviral therapeutics or vaccines are lacking. In this study, we investigated the role of host-cell chloride (Cl-) channels on CHIKV replication.We demonstrate that specific pharmacological Cl- channel inhibitors significantly inhibit CHIKV replication in a dose-dependent manner, suggesting that Cl-channels are pro-viral factors in human cells. Further analysis of the effect of the inhibitors on CHIKV attachment, entry, viral protein expression and replicon replication demonstrated that Cl- channels are specifically required for efficient CHIKV genome replication. This was conserved in mosquito cells, where CHIKV replication and genome copy number was significantly reduced following Cl- channel inhibition. siRNA silencing identified chloride intracellular channels 1 and 4 (CLIC1 and CLIC4, respectively) as required for efficient CHIKV replication and protein affinity chromatography showed low levels of CLIC1 in complex with CHIKV nsP3, an essential component of the viral replication machinery. In summary, for the first time we demonstrate that efficient replication of the CHIKV genome depends on cellular Cl- channels, in both human and mosquito cells and identifies CLIC1 and CLIC4 as agonists of CHIKV replication in human cells. We observe a modest interaction, either direct or indirect, between CLIC1 and nsP3 and hypothesize that CLIC1 may play a role in the formation/maintenance of CHIKV replication complexes. These findings advance our molecular understanding of CHIKV replication and identify potential druggable targets for the treatment and prevention of CHIKV mediated disease.

Comprehensive analysis of the host-virus interactome of SARS-CoV-2

“…When ORF3a was overexpressed, the localization of CLCC1 changed to strong co-localization with ORF3a in the endosome and lysosome. Early reports suggest that chloride channel function may be required for the viral lifecycle 13,14 . Our results indicate that SARS-CoV-2 may take over the function of CLCC1 protein to promote its own lifecycle.…”

TMEM16A/ANO1 calcium-activated chloride channel as a novel target for the treatment of human respiratory syncytial virus infection

“…The organisation, composition and functions of replication factories induced by RNA viruses are generally accepted to require the recruitment of a range of host cell factors. 43 Anion channels are important in a range of these viral replicase systems; CHIKV is dependent on CLIC1 that interacts with NSP3, a known component of the replicase complex 12 ; the voltage-dependent anion channel 1 directly interacts with VP1 and VP3 of infectious bursal disease virus, a member of the Birnaviridae to stabilise RNP formation 44 ; and CLIC-1, ClC-5 and ClC-7 silencing impede HCV replication. 13 Of interest, inclusion body-associated granules (IBAGs) are key to HRSV RNA synthesis and contain viral mRNA and the viral protein M2-1, but exclude viral genomes and all necessary components required for transcription and replication (N, P and L).…”

Persistent Joint Pain Following Arthropod Virus Infections

“…CHIKV enters and replicates in a variety of cell lineages, including fibroblasts and macrophages at the infection site. It can then travel systemically through the lymph nodes, spleen, liver, muscles, and joints [ 59 – 62 ]. MAYV also targets macrophages in order to replicate [ 63 ].…”