Murine cytomegalovirus open reading frame m29.1 augments virus replication both in vitro and in vivo.
Murine cytomegalovirus mutant Rc29, with a premature stop codon mutation in the m29 open reading frame (ORF), produced no apparent phenotype in cell culture or following infection of BALB/c mice. In contrast, a similar mutant virus, Rc29.1, with a premature stop codon mutation in its m29.1 ORF, showed reduced virus yields (2-3 log(10) p.f.u. ml(-1)) in tissue culture. Mutant virus yields in BALB/c mice were delayed, reduced ( approximately 1 log(10) p.f.u. per tissue) and persisted less well in salivary glands compared with wild-type (wt) and revertant (Rv29.1) virus. In severe combined immunodeficiency mice, Rc29.1 virus showed delayed and reduced replication initially in all tissues (liver, spleen, kidneys, heart, lung and salivary glands). This delayed death until 31 days post-infection (p.i.) compared with wt (23 days p.i.) but at death virus yields were similar to wt. m29 gene transcription was initiated at early times post-infection, while production of a transcript from ORF m29.1 in the presence of cycloheximide indicated that it was an immediate-early gene. ORFs m29.1 and M28 are expressed from a bicistronic message, which is spliced infrequently. However, it is likely that each ORF expresses its own protein, as antiserum derived in rabbits to the m29.1 protein expressed in bacteria from the m29.1 ORF detected only one protein in Western blot analysis of the size predicted for the m29.1 protein. Our results suggest that neither ORF is essential for virus replication but m29.1 is important for optimal viral growth in vitro and in vivo.
Ahasan
Mohammad M
MM
School of Biosciences, The University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
Sweet
Clive
C
School of Biosciences, The University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
D000595:Amino Acid Sequence; D000825:Animal Structures / Q000821:virology; D000818:Animals; D000956:Antigens, Viral / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D001483:Base Sequence; D018389:Codon, Nonsense; D006566:Herpesviridae Infections / Q000821:virology; D017874:Immediate-Early Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D016513:Mice, SCID; D008969:Molecular Sequence Data; D018146:Muromegalovirus / Q000235:genetics / Q000502:physiology*; D016296:Mutagenesis; D041681:NIH 3T3 Cells; D016366:Open Reading Frames; D017354:Point Mutation; D016019:Survival Analysis; D013997:Time Factors; D010948:Viral Plaque Assay; D014779:Virus Replication
D000956:Antigens, Viral; D018389:Codon, Nonsense; D017874:Immediate-Early Proteins; C044582:immediate-early proteins, cytomegalovirus