title: UL54-null pseudorabies virus is attenuated in mice but productively infects cells in culture.
abstract: The pseudorabies virus (PRV) UL54 homologs are important multifunctional proteins with roles in shutoff of host protein synthesis, transactivation of virus and cellular genes, and regulation of splicing and translation. Here we describe the first genetic characterization of UL54. We constructed UL54 null mutations in a PRV bacterial artificial chromosome using sugar suicide and lambdaRed allele exchange systems. Surprisingly, UL54 is dispensable for growth in tissue culture but exhibits a small-plaque phenotype that can be complemented in trans by both the herpes simplex virus type 1 ICP27 and varicella-zoster virus open reading frame 4 proteins. Deletion of UL54 in the virus vJSdelta54 had no effect on the ability of the virus to shut off host cell protein synthesis but did affect virus gene expression. The glycoprotein gC accumulated to lower levels in cells infected with vJSdelta54 compared to those infected with wild-type virus, while gK levels were undetectable. Other late gene products, gB, gE, and Us9, accumulated to higher levels than those seen in cells infected with wild-type virus in a multiplicity-dependent manner. DNA replication is also reduced in cells infected with vJSdelta54. UL54 appears to regulate UL53 and UL52 at the transcriptional level as their respective RNAs are decreased in cells infected with vJSdelta54. Interestingly, vJSdelta54 is highly attenuated in a mouse model of PRV infection. Animals infected with vJSdelta54 survive twice as long as animals infected with wild-type virus, and this results in delayed accumulation of virus-specific antigens in skin, dorsal root ganglia, and spinal cord tissues.
authors.affiliation: Department of Microbiology, Columbia University, 701 W. 168th St., New York, NY 10032, USA.
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Schwartz
authors.forename: Elizabeth E
authors.initials: EE
authors.lastname: Brittle
authors.forename: Ashley E
authors.initials: AE
authors.lastname: Reynolds
authors.forename: Lynn W
authors.initials: LW
authors.lastname: Enquist
authors.forename: Saul J
authors.initials: SJ
authors.lastname: Silverstein
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000378:metabolism; D002460:Cell Line; D004195:Disease Models, Animal; D005727:Ganglia, Spinal / Q000378:metabolism; D015870:Gene Expression; D005786:Gene Expression Regulation; D005814:Genes, Viral / Q000235:genetics; D005816:Genetic Complementation Test; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D017874:Immediate-Early Proteins / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D011557:Pseudorabies / Q000821:virology*; D012867:Skin / Q000378:metabolism; D013116:Spinal Cord / Q000378:metabolism; D013997:Time Factors; D015534:Trans-Activators; D014764:Viral Proteins / Q000235:genetics; D014779:Virus Replication
chemical_list: D000956:Antigens, Viral; D006023:Glycoproteins; C057120:ICP27 protein, human herpesvirus 1; D017874:Immediate-Early Proteins; C085824:ORF4 protein, Human herpesvirus 3; D015534:Trans-Activators; D014764:Viral Proteins