Reduced Levels of IE2 Gene Expression and Shutdown of Early and Late Viral Genes during Latent Infection of the Glioblastoma Cell Line U138-MG with Selectable Recombinants of Human Cytomegalovirus

To establish stable culture conditions which support persistence of the human cytomegalovirus (HCMV) genome in a latent state, the expression of the bacterial neomycin phosphotransferase (neo) from HCMV recombinants was used for selection. Different cell lines were infected with HCMV recombinants. T...

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Veröffentlicht in:Virology (New York, N.Y.) N.Y.), 1994-10, Vol.204 (1), p.101-113
Hauptverfasser: Wolff, Dietmar, Sinzger, Christian, Drescher, Petra, Jahn, Gerhard, Plachter, Bodo
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container_issue 1
container_start_page 101
container_title Virology (New York, N.Y.)
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creator Wolff, Dietmar
Sinzger, Christian
Drescher, Petra
Jahn, Gerhard
Plachter, Bodo
description To establish stable culture conditions which support persistence of the human cytomegalovirus (HCMV) genome in a latent state, the expression of the bacterial neomycin phosphotransferase (neo) from HCMV recombinants was used for selection. Different cell lines were infected with HCMV recombinants. The human glioblastoma line U138-MG was rendered resistant to G418 and retained the vital genome. More than 90% of the cells expressed the vital IE1 protein of 72 kDa for a culture period of 18 months. Many fewer cells expressed IE2-encoded proteins. No late gene expression or infectious virus was detectable. IE2 gene expression in latently infected cells appeared to be restricted at the level of RNA accumulation. Treatment with TPA or retinoic acid led to enhanced expression of the IE2 gene and the early genes encoding pp65 (UL83) and p52 (UL44). Superinfection with wild-type HCMV led to replication of neo-recombinant virus, indicating that replication-competent virus had been retained in latently infected U138-MG and that the cells had kept their permissive phenotype. Latent HCMV infection in U138-MG cells provides a useful model system for studying the role of particular viral and cellular genes in latent and permissive infections.
doi_str_mv 10.1006/viro.1994.1514
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subjects Cytomegalovirus - genetics
Cytomegalovirus - physiology
DNA, Viral - analysis
Gene Expression Regulation, Viral - genetics
Genes, Immediate-Early - genetics
Genome, Viral
Glioblastoma - microbiology
Humans
Immediate-Early Proteins - genetics
Kanamycin Kinase
Membrane Glycoproteins
Phosphotransferases (Alcohol Group Acceptor) - genetics
RNA, Messenger - analysis
RNA, Viral - analysis
Trans-Activators
Tumor Cells, Cultured
Viral Envelope Proteins
Viral Proteins - analysis
Virus Activation
Virus Latency - genetics
Virus Replication
title Reduced Levels of IE2 Gene Expression and Shutdown of Early and Late Viral Genes during Latent Infection of the Glioblastoma Cell Line U138-MG with Selectable Recombinants of Human Cytomegalovirus
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