Autographa californica M nuclear polyhedrosis virus: Microtubules and replication

Abstract

Progressive reorganization and depolymerization of microtubules corresponded with virus-induced rounding of Autographa californica M nuclear polyhedrosis virus (AcMNPV)-infected Spodoptera frugiperda IPLB-Sf-21 cells, suggesting that microtubules were instrumental in maintaining the normal shape of these cells. Depolymerization of all cortical and most of the paranuclear microtubules with colchicine also resulted in cell rounding, confirming this hypothesis. Studies with aphidicolin and cycloheximide indicated the virus-induced effects on the microtubules were mediated by both early and late viral gene products. Microtubules in cells infected with a p10 deletion mutant depolymerized microtubules in a manner similar to those in wild-type virus-infected cells, indicating p10 was not responsible for virus-induced changes in the microtubules. Nevertheless, evidence for the association of p10 and microtubules was obtained by fluorescence microscopy and immunoelectron microscopy. Colchicine depolymerization of microtubules before and throughout infection did not interfere with virus replication, but treatment of cells with taxol, a microtubule-stabilizing agent, both delayed and depressed virus replication. The taxol-induced effect was relieved by the addition of colchicine. These results suggested that AcMNPV-induced depolymerization of microtubules may be a necessary event in, rather than a tangential effect of, virus replication. Attempts to monitor the effects of virus infection on intermediate filaments were unsuccessful due to the lack of cross-reactivity between antibodies to intermediate filament proteins and IPLBSf-21 cells, indicating these proteins are not highly conserved in lepidopteran insect cells.