title: Transcriptional Silencing of Moloney Murine Leukemia Virus in Human Embryonic Carcinoma Cells.
abstract: Embryonic carcinoma (EC) cells are malignant counterparts of embryonic stem (ES) cells and serve as useful models for investigating cellular differentiation and human embryogenesis. Though the susceptibility of murine EC cells to retroviral infection has been extensively analyzed, few studies of retrovirus infection of human EC cells have been performed. We tested the susceptibility of human EC cells to transduction by retroviral vectors derived from three different retroviral genera. We show that human EC cells efficiently express reporter genes delivered by vectors based on human immunodeficiency virus type 1 (HIV-1) and Mason-Pfizer monkey virus (M-PMV) but not Moloney murine leukemia virus (MLV). In human EC cells, MLV integration occurs normally, but no viral gene expression is observed. The block to MLV expression of MLV genomes is relieved upon cellular differentiation. The lack of gene expression is correlated with transcriptional silencing of the MLV promoter through the deposition of repressive histone marks as well as DNA methylation. Moreover, depletion of SETDB1, a histone methyltransferase, resulted in a loss of transcriptional silencing and upregulation of MLV gene expression. Finally, we provide evidence showing that the lack of MLV gene expression may be attributed in part to the lack of MLV enhancer function in human EC cells.
  
  
  OBJECTIVE
  Human embryonic carcinoma (EC) cells are shown to restrict the expression of murine leukemia virus genomes but not retroviral genomes of the lentiviral or betaretroviral families. The block occurs at the level of transcription and is accompanied by the deposition of repressive histone marks and methylation of the integrated proviral DNA. The host machinery required for silencing in human EC cells is distinct from that in murine EC cell lines: the histone methyltransferase SETDB1 is required, but the widely utilized corepressor TRIM28/Kap1 is not. A transcriptional enhancer element from the Mason-Pfizer monkey virus can override the silencing and promote transcription of chimeric proviral DNAs. The findings reveal novel features of human EC gene regulation not present in their murine counterparts.
authors: Integrated Program in Cellular, Molecular and Biophysical Studies, Columbia University, New York, New York, USA.|Medical Scientist Training Program, Columbia University College of Physicians and Surgeons, New York, New York, USA.
authors: Gary Z
authors: GZ
authors: Wang
authors: Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York, USA spg1@cumc.columbia.edu.|Department of Microbiology and Immunology, Columbia University, New York, New York, USA.|Howard Hughes Medical Institute, Columbia University, New York, New York, USA.
authors: Stephen P
authors: SP
authors: Goff
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D019175:DNA Methylation; D020868:Gene Silencing; D017930:Genes, Reporter; D016679:Genome, Viral; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism; D011495:Histone-Lysine N-Methyltransferase; D006657:Histones / Q000235:genetics / Q000276:immunology; D058507:Host Specificity; D000066449:Human Embryonic Stem Cells / Q000276:immunology* / Q000821:virology; D006801:Humans; D016093:Mason-Pfizer monkey virus / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008979:Moloney murine leukemia virus / Q000235:genetics* / Q000378:metabolism; D014411:Neoplastic Stem Cells / Q000276:immunology* / Q000821:virology; D011401:Promoter Regions, Genetic; D011496:Protein Methyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D013045:Species Specificity; D014158:Transcription, Genetic
chemical_list: D006657:Histones; D034741:RNA, Small Interfering; D011496:Protein Methyltransferases; D011495:Histone-Lysine N-Methyltransferase; C454062:SETDB1 protein, human
keywords: DNA methylation; chromatin immunoprecipitation; enhancer; histones; repressor