Wolbachia interferes with Zika virus replication by hijacking cholesterol metabolism in mosquito cells.

Zika virus is a member of the arbovirus family transmitted by mosquitos and it is associated with microcephaly in infants born to infected mothers. is an intracellular gram-negative alpha-proteobacteria that infects many species of arthropods, including mosquitos. The presence of in mosquitos has been shown to control the vector population and suppress arbovirus transmission. One mechanism of -mediated interference with virus replication is competition over host resources between and the virus. We hypothesize that cholesterol metabolism is involved in -mediated virus suppression due to its important role in Zika virus replication. In this study, we determined that impacted virus replication by altering cholesterol biosynthesis in C6/36 cells, diverting resources from the host cell mevalonate (MVA) pathway to fulfill the needs of the bacteria. This resulted in a decrease of total cholesterol, increased loads, and decreased viral titers. Inhibition of the MVA pathway using fluvastatin decreased total cholesterol and viral titers, mimicking the effects of on the virus in free cells. We also found that infected cells had depleted lipid droplets, the main component of which is cholesterol esters. We confirmed that cholesterol esterases were upregulated in response to virus infection in C6/36 cells. Functional analysis showed that alteration of cholesterol metabolism simulated -mediated inhibition of virus infection in C6/36 cells. Our study provides a mechanism behind -induced interference of arbovirus replication and could help advance strategies to control arbovirus pathogens in insect vectors and human infections. IMPORTANCE Arthropod-borne viruses are emerging pathogens that are spread widely by mosquitos. Zika virus is an arbovirus that can infect humans and be transmitted from an infected mother to the fetus, potentially leading to microcephaly in infants. One promising strategy to prevent disease caused by arboviruses is to target the insect vector population. Recent field studies have shown that mosquito populations infected with bacteria suppress arbovirus replication and transmission. Here, we describe how intracellular bacteria redirect resources within their host cells and suppress Zika virus replication at the cellular level. Understanding the mechanism behind -induced interference of arbovirus replication could help advance strategies to control arbovirus pathogens in insect vectors and human populations.