Kinesin-1 transports morphologically distinct intracellular virions during vaccinia infection

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Abstract

Intracellular mature virus (IMV) are the first and most abundant infectious form of vaccinia virus to assemble during its replication cycle. IMV can undergo microtubule-based motility, but their directionality and the motor involved in their transport remain unknown. Here, we demonstrate that IMV, like intracellular enveloped virus (IEV), the second form of vaccinia that is wrapped in Golgi-derived membrane, recruit kinesin-1 and undergo anterograde transport. In vitro reconstitution of virion transport in infected cell extracts reveals that IMV and IEV move toward microtubule plus-ends with respective velocities of 0.66 and 0.56 µm/s. Quantitative imaging establishes that IMV and IEV recruit an average of 139 and 320 kinesin-1 motor complexes respectively. In the absence of kinesin-1 there is a near-complete loss of in vitro motility and reduction in the intracellular spread of both types of virion. Our observations demonstrate that kinesin-1 transports two morphologically distinct forms of vaccinia. Reconstitution of vaccinia-based microtubule motility in vitro provides a new model to elucidate how motor number and regulation impacts transport of a bona fide kinesin-1 cargo.