title: Complex expression of the UL136 gene of human cytomegalovirus results in multiple protein isoforms with unique roles in replication.
abstract: Human cytomegalovirus (HCMV) is a complex DNA virus with a 230-kb genome encoding 170 and up to 750 proteins. The upper limit of this coding capacity suggests the evolution of complex mechanisms to substantially increase the coding potential from the 230-kb genome. Our work examines the complexity of one gene, UL136, encoded within the ULb' region of the genome that is lost during serial passage of HCMV in cultured fibroblasts. UL136 is expressed as five protein isoforms. We mapped these isoforms and demonstrate that they originate from both a complex transcriptional profile and, possibly, the usage of multiple translation initiation sites. Intriguingly, the pUL136 isoforms exhibited distinct subcellular distributions with varying association with the Golgi apparatus. The subcellular localization of membrane-bound isoforms of UL136 differed between when they were expressed exogenously and when they were expressed in the context of viral infection, suggesting that the trafficking of these isoforms is mediated by infection-specific factors. While UL136, like most ULb' genes, was dispensable for replication in fibroblasts, the soluble 23- and 19-kDa isoforms suppressed virus replication. In CD34(+) hematopoietic progenitor cells (HPCs) infected in vitro, disruption of the 23- and 19-kDa isoforms resulted in increased replication and a loss of the latency phenotype, similar to the effects of the UL138 latency determinant encoded within the same genetic locus. Our work suggests a complex interplay between the UL136 isoforms which balances viral replication in multiple cell types and likely contributes to the cell type-dependent phenotypes of the UL133/8 locus and the outcome of HCMV infection.
  
  
  OBJECTIVE
  HCMV is a significant cause of morbidity in immunocompromised individuals, including transplant patients. The lifelong persistence of the virus results in a high seroprevalence worldwide and may contribute to age-related pathologies, such as atherosclerosis. The mechanisms of viral persistence are poorly understood; however, understanding the molecular basis of persistence is imperative for the development of new treatments. In this work, we characterize a complex HCMV gene, UL136, which is expressed as five protein isoforms. These isoforms arise predominantly from complex transcriptional mechanisms, which contribute to an increased coding capacity of the virus. Further, the UL136 isoforms oppose the activity of one another to balance HCMV replication in multiple cell types. We identify soluble isoforms of UL136 that function to suppress virus replication in fibroblasts and in CD34(+) HPCs for latency.
authors.affiliation: Graduate Interdisciplinary Program in Genetics, University of Arizona, Tucson, Arizona, USA.
authors.forename: Katie
authors.initials: K
authors.lastname: Caviness
authors.affiliation: BIO5 Institute, University of Arizona, Tucson, Arizona, USA.
authors.forename: Louis
authors.initials: L
authors.lastname: Cicchini
authors.affiliation: Department of Cellular and Molecular Medicine, University of Arizona, Tucson, Arizona, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Rak
authors.affiliation: BIO5 Institute, University of Arizona, Tucson, Arizona, USA.
authors.forename: Mahadevaiah
authors.initials: M
authors.lastname: Umashankar
authors.affiliation: Graduate Interdisciplinary Program in Genetics, University of Arizona, Tucson, Arizona, USA BIO5 Institute, University of Arizona, Tucson, Arizona, USA Department of Cellular and Molecular Medicine, University of Arizona, Tucson, Arizona, USA Department of Immunobiology, University of Arizona, Tucson, Arizona, USA fgoodrum@email.arizona.edu.
authors.forename: Felicia
authors.initials: F
authors.lastname: Goodrum
mesh_terms: D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000502:physiology*; D005347:Fibroblasts / Q000821:virology; D017353:Gene Deletion; D020869:Gene Expression Profiling; D006412:Hematopoietic Stem Cells / Q000821:virology; D006801:Humans; D015388:Organelles / Q000737:chemistry; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D020033:Protein Isoforms; D014764:Viral Proteins