bioRxiv Science⌕ Search

Biology subjects

Gil, P. I.

Publications and source records attributed to Gil, P. I..

2 recordsLinked to original sources

Actin cytoskeleton dynamics affect replication of Human Metapneumovirus

Human Metapneumovirus (hMPV) is responsible for viral respiratory infection with clinical and epidemiological relevance in pediatric, immunocompromised, and elderly populations. Little is known about hMPV in vitro replication processes and their relationship with cellular structures such as the cytoskeleton. Our goal was to evaluate the role of the actin cytoskeleton in hMPV replication at different stages of viral growth. hMPV was isolated in Vero cells from a clinical sample and identified as A2 genotype. The cytopathic effect was detected by the appearance of cell rounding and refractory cell clusters. The growth curve showed that viral replication maximum level was between 48 and 72 hpi. The highest percentage of infected cells and intracellular hMPV-protein were detected at the early stages of the replication cycle. Disruption of actin microfilaments with Cytochalasin D during the early events provoked an increase in both intracellular and extracellular viruses. We demonstrate that the early phase of the hMPV curve is crucial for viral replication; and the disruption of microfilaments during this time increments both viral protein expression and release of viruses to the extracellular space. This study contributes to elucidate wild-type hMPV growth kinetics, providing new insights on the actin cytoskeleton role in viral replication mechanisms.

microbiology↗

Functional entry of Members of Venezuelan Equine Encephalitis Complex into host cells is dependent on clathrin-mediated endocytosis, in a pH-Dependent Manner

Pixuna virus (PIXV) and Rio Negro virus (RNV) are mosquito-borne alphaviruses belonging to the Venezuelan Equine Encephalitis (VEE) complex, which includes pathogenic epizootic and enzootic subtypes responsible for life-threatening diseases in equines. Considering that the first steps in viral infection are crucial for the efficient production of new progeny, the aim of this study was to elucidate the early events of the replication cycle of these two viruses. To this end, we used chemical inhibitors and the expression of dominant-negative constructs to study the dependence of clathrin and endosomal pH on PIXV and RNV internalization mechanisms. We demonstrated that both viruses are internalized primarily via clathrin-mediated endocytosis, where the low pH in endosomes is crucial for viral replication. Contributing knowledge regarding the entry route of VEE complex members is important to understand the pathogenesis of these viruses and also to develop new antiviral strategies.

microbiology↗