Abstract
The role of the intranuclear movement of chromatin in gene expression is not well-understood. Herpes simplex virus forms replication compartments (RCs) in infected cell nuclei as sites of viral DNA replication and late gene transcription. These structures develop from small compartments that grow in size, move, and coalesce. Quantitative analysis of RC trajectories, derived from 4D images, shows that most RCs move by directed motion. Directed movement is impaired in the presence of actin and myosin inhibitors as well as a transcription inhibitor. In addition, RCs coalesce at and reorganize nuclear speckles. Lastly, distinct effects of actin and myosin inhibitors on viral gene expression suggest that RC movement is not required for transcription, but rather, movement results in the bridging of transcriptionally active RCs with nuclear speckles to form structures that enhance export of viral late mRNAs.
Highlights
The role of the intranuclear movement of chromatin in gene expression is not well-understood
Other studies have shown that herpes simplex virus (HSV) capsids undergo directed movement inside the nucleus [21], which is inhibited by the actin polymerization inhibitor latrunculin A and the putative myosin inhibitor 2,3-butanedione monoxime (BDM) [22, 23]
replication compartments (RCs), which were first identified by the presence of infected cell protein (ICP) 8 [15], develop from small structures that grow in size, move, and coalesce, filling the entire nucleus and marginalizing host chromatin to the nuclear periphery [17, 25,26,27]
Summary
The role of the intranuclear movement of chromatin in gene expression is not well-understood. To better understand the mechanism and function of intranuclear movement of RCs, we carried out 4D imaging of cells infected with a recombinant HSV-1 strain expressing ICP8 fused to GFP to visualize RCs. Our results showed that the majority of RCs move by directed motion and require nuclear actin, myosin, and ongoing transcription. To determine the role of nuclear actin in RC movement, we treated cells with lat A at 3 hpi.
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