Viral Mimicry of Cdc2/Cyclin-Dependent Kinase 1 Mediates Disruption of Nuclear Lamina during Human Cytomegalovirus Nuclear Egress

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Abstract
The nuclear lamina is a major obstacle encountered by herpesvirus nucleocapsids in their passage from the nucleus to the cytoplasm (nuclear egress). We found that the human cytomegalovirus (HCMV)-encoded protein kinase UL97, which is required for efficient nuclear egress, phosphorylates the nuclear lamina component lamin A/C in vitro on sites targeted by Cdc2/cyclin-dependent kinase 1, the enzyme that is responsible for breaking down the nuclear lamina during mitosis. Quantitative mass spectrometry analyses, comparing lamin A/C isolated from cells infected with viruses either expressing or lacking UL97 activity, revealed UL97-dependent phosphorylation of lamin A/C on the serine at residue 22 (Ser22). Transient treatment of HCMV-infected cells with maribavir, an inhibitor of UL97 kinase activity, reduced lamin A/C phosphorylation by approximately 50%, consistent with UL97 directly phosphorylating lamin A/C during HCMV replication. Phosphorylation of lamin A/C during viral replication was accompanied by changes in the shape of the nucleus, as well as thinning, invaginations, and discrete breaks in the nuclear lamina, all of which required UL97 activity. As Ser22 is a phosphorylation site of particularly strong relevance for lamin A/C disassembly, our data support a model wherein viral mimicry of a mitotic host cell kinase activity promotes nuclear egress while accommodating viral arrest of the cell cycle. Human cytomegalovirus (HCMV) causes life-threatening disease in transplant patients and people with AIDS, and is also an important cause of birth defects. Like all viruses, HCMV must have a way to leave the host cell, so that it can infect new cells. Moreover, as a member of the herpesvirus family, HCMV replicates its DNA in the nucleus, so it must have mechanisms to ensure that its genetic material can exit from the nucleus (nuclear egress). HCMV encodes a protein kinase, UL97, which is required for efficient nuclear egress. We found that UL97 aids nuclear egress by mimicking a host cell enzyme that normally helps break down a protein meshwork in the nucleus during cell division. The enzyme activity of UL97 pokes holes in the meshwork that allow nascent HCMV virions to gain access to the nuclear membrane. UL97 is also an important target for drugs for treating HCMV disease. This work not only helps explain how these drugs act, but also highlights the potential of targeting nuclear egress for the discovery of new drugs.