Viral mimicry of Cdc2/cyclin-dependent kinase 1 mediates disruption of nuclear lamina during human cytomegalovirus nuclear egress.
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 (Ser(22)). 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 Ser(22) 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.
Hamirally
Sofia
S
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, United States of America.
Kamil
Jeremy P
JP
Ndassa-Colday
Yasmine M
YM
Lin
Alison J
AJ
Jahng
Wan Jin
WJ
Baek
Moon-Chang
MC
Noton
Sarah
S
Silva
Laurie A
LA
Simpson-Holley
Martha
M
Knipe
David M
DM
Golan
David E
DE
Marto
Jarrod A
JA
Coen
Donald M
DM
D001562:Benzimidazoles / Q000494:pharmacology; D016203:CDC2 Protein Kinase / Q000235:genetics*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003587:Cytomegalovirus / Q000502:physiology*; D003586:Cytomegalovirus Infections / Q000503:physiopathology; D006801:Humans; D034904:Lamin Type A / Q000378:metabolism; D018716:Molecular Mimicry / Q000502:physiology*; D034881:Nuclear Lamina / Q000378:metabolism; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012263:Ribonucleosides / Q000494:pharmacology; D014779:Virus Replication / Q000502:physiology; D004004:Dichlororibofuranosylbenzimidazole / Q000031:analogs & derivatives
D001562:Benzimidazoles; D016203:CDC2 Protein Kinase; D034904:Lamin Type A; D017853:Phosphotransferases (Alcohol Group Acceptor); D012263:Ribonucleosides; C494847:LMNA protein, human; C075365:ganciclovir kinase; C400401:maribavir; D004004:Dichlororibofuranosylbenzimidazole