Control of cytoplasmic maturation events by cytomegalovirus tegument protein pp150.
Cytomegalovirus replication depends upon a betaherpesvirus-conserved 150-kDa tegument phosphoprotein (pp150; encoded by UL32) that supports the final steps in virion maturation at cytoplasmic assembly compartments. Amino acid substitutions were introduced into conserved region 1 (CR1) and CR2 of pp150, affecting a region that may interact with nucleocapsids. Two independent CR2 point mutants (N201A and G207A) failed to support viral replication in evaluations by a transient complementation assay or after reconstruction into recombinant viruses. An assembly compartment-like cytoplasmic inclusion developed in UL32 mutant virus-infected cells that was similar to that of wild-type virus-infected cells. The cellular localization of the trans-Golgi marker Golgin-97 suggested differences in the organization of the assembly compartment compared to that of wild-type virus-infected cells. Replication-defective CR2 point mutants exhibited the same phenotype as that of a virus carrying a complete deletion of the UL32 open reading frame in these assays. Electron micrographs of fibroblasts at 3 or 5 days postinfection with a deletion mutant (DeltaUL32) grown on UL32-complementing cells showed a similar number and morphology of capsids in the nucleus, but the cytoplasmic region associated with virion assembly appeared highly vesiculated and contained few recognizable nucleocapsids or complete virus particles. These data demonstrate that the principle role of pp150 is to retain nucleocapsid organization through secondary envelopment at the assembly compartment.
Tandon
Ritesh
R
Emory Vaccine Center, 1462 Clifton Road, Atlanta, GA 30322, USA.
Mocarski
Edward S
ES
D000595:Amino Acid Sequence; D001324:Autoantigens / Q000378:metabolism; D022202:Chromosomes, Artificial, Bacterial; D003587:Cytomegalovirus / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D000077614:Golgi Matrix Proteins; D006801:Humans; D008954:Models, Biological; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins / Q000737:chemistry; D016366:Open Reading Frames; D010750:Phosphoproteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D017386:Sequence Homology, Amino Acid; D014763:Viral Matrix Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D014764:Viral Proteins / Q000378:metabolism*
D001324:Autoantigens; D000077614:Golgi Matrix Proteins; C450721:Golgi complex autoantigen, 97-kDa; D019590:Nucleocapsid Proteins; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; D014764:Viral Proteins; C056367:pp150 protein, Cytomegalovirus