title: Inactivation of retinoblastoma protein does not overcome the requirement for human cytomegalovirus UL97 in lamina disruption and nuclear egress.
abstract: Human cytomegalovirus (HCMV) encodes one conventional protein kinase, UL97. During infection, UL97 phosphorylates the retinoblastoma tumor suppressor protein (pRb) on sites ordinarily phosphorylated by cyclin-dependent kinases (CDK), inactivating the ability of pRb to repress host genes required for cell cycle progression to S phase. UL97 is important for viral DNA synthesis in quiescent cells, but this function can be replaced by human papillomavirus type 16 E7, which targets pRb for degradation. However, viruses in which E7 replaces UL97 are still defective for virus production. UL97 is also required for efficient nuclear egress of viral nucleocapsids, which is associated with disruption of the nuclear lamina during infection, and phosphorylation of lamin A/C on serine 22, which antagonizes lamin polymerization. We investigated whether inactivation of pRb might overcome the requirement of UL97 for these roles, as pRb inactivation induces CDK1, and CDK1 phosphorylates lamin A/C on serine 22. We found that lamin A/C serine 22 phosphorylation during HCMV infection correlated with expression of UL97 and was considerably delayed in UL97-null mutants, even when E7 was expressed. E7 failed to restore gaps in the nuclear lamina seen in wild-type but not UL97-null virus infections. In electron microscopy analyses, a UL97-null virus expressing E7 was as impaired as a UL97-null mutant in cytoplasmic accumulation of viral nucleocapsids. Our results demonstrate that pRb inactivation is insufficient to restore efficient viral nuclear egress of HCMV in the absence of UL97 and instead argue further for a direct role of UL97 in this stage of the infectious cycle.
authors: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, USA.
authors: Natalia I
authors: NI
authors: Reim
authors: Jeremy P
authors: JP
authors: Kamil
authors: Depeng
authors: D
authors: Wang
authors: Alison
authors: A
authors: Lin
authors: Mayuri
authors: M
authors: Sharma
authors: Maria
authors: M
authors: Ericsson
authors: Jean M
authors: JM
authors: Pesola
authors: David E
authors: DE
authors: Golan
authors: Donald M
authors: DM
authors: Coen
mesh_terms: D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism / Q000821:virology; D003587:Cytomegalovirus / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D003586:Cytomegalovirus Infections / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D006801:Humans; D034904:Lamin Type A / Q000737:chemistry / Q000378:metabolism; D034881:Nuclear Lamina / Q000737:chemistry / Q000378:metabolism / Q000821:virology*; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000378:metabolism*; D058105:Polymerization; D016160:Retinoblastoma Protein / Q000235:genetics / Q000378:metabolism*; D057074:Virus Release
chemical_list: D034904:Lamin Type A; D016160:Retinoblastoma Protein; C440308:lamin C; D017853:Phosphotransferases (Alcohol Group Acceptor); C075365:ganciclovir kinase