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dc.contributor.authorLawrence, Jeanne B.
dc.contributor.authorMarselle, Lisa M.
dc.contributor.authorByron, Kevin S.
dc.contributor.authorJohnson, Carol V.
dc.contributor.authorSullivan, John L.
dc.contributor.authorSinger, Robert H.
dc.date2022-08-11T08:09:36.000
dc.date.accessioned2022-08-23T16:37:23Z
dc.date.available2022-08-23T16:37:23Z
dc.date.issued1990-07-01
dc.date.submitted2009-04-02
dc.identifier.citationProc Natl Acad Sci U S A. 1990 Jul;87(14):5420-4. <a href="http://www.pnas.org/content/87/14/5420.full.pdf+html">Link to article on publisher's website</a>
dc.identifier.issn0027-8424 (Print)
dc.identifier.pmid2371279
dc.identifier.urihttp://hdl.handle.net/20.500.14038/39005
dc.description.abstractDetection and subcellular localization of human immunodeficiency virus (HIV) were investigated using sensitive high-resolution in situ hybridization methodology. Lymphocytes infected with HIV in vitro or in vivo were detected by fluorescence after hybridization with either biotin or digoxigenin-labeled probes. At 12 hr after infection in vitro, a single intense signal appeared in the nuclei of individual cells. Later in infection, when cytoplasmic fluorescence became intense, multiple nuclear foci frequently appeared. The nuclear focus consisted of newly synthesized HIV RNA as shown by hybridization in the absence of denaturation and by susceptibility to RNase and actinomycin D. Virus was detected in patient lymphocytes and it was shown that a singular nuclear focus also characterizes cells infected in vivo. The cell line 8E5/LAV containing one defective integrated provirus revealed a similar focus of nuclear RNA, and the single integrated HIV genome was unequivocally visualized on a D-group chromosome. This demonstrates an extremely sensitive single-cell assay for the presence of a single site of HIV transcription in vitro and in vivo and suggests that it derives from one (or very few) viral genomes per cell. In contrast, productive Epstein-Barr virus infection exhibited many foci of nuclear RNA per cell.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=2371279&dopt=Abstract">Link to Article in PubMed</a>
dc.relation.urlhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC54336/pdf/pnas01039-0196.pdf
dc.subjectCell Nucleus
dc.subjectCells, Cultured
dc.subjectDactinomycin
dc.subject*Genes, Viral
dc.subjectHIV-1
dc.subjectHumans
dc.subjectKinetics
dc.subjectMicroscopy, Fluorescence
dc.subjectNucleic Acid Hybridization
dc.subjectRNA, Viral
dc.subjectRibonuclease, Pancreatic
dc.subjectT-Lymphocytes
dc.subjectTranscription, Genetic
dc.subjectCell Biology
dc.subjectLife Sciences
dc.subjectMedicine and Health Sciences
dc.titleSubcellular localization of low-abundance human immunodeficiency virus nucleic acid sequences visualized by fluorescence in situ hybridization
dc.typeJournal Article
dc.source.journaltitleProceedings of the National Academy of Sciences of the United States of America
dc.source.volume87
dc.source.issue14
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/oapubs/1830
dc.identifier.contextkey808596
html.description.abstract<p>Detection and subcellular localization of human immunodeficiency virus (HIV) were investigated using sensitive high-resolution in situ hybridization methodology. Lymphocytes infected with HIV in vitro or in vivo were detected by fluorescence after hybridization with either biotin or digoxigenin-labeled probes. At 12 hr after infection in vitro, a single intense signal appeared in the nuclei of individual cells. Later in infection, when cytoplasmic fluorescence became intense, multiple nuclear foci frequently appeared. The nuclear focus consisted of newly synthesized HIV RNA as shown by hybridization in the absence of denaturation and by susceptibility to RNase and actinomycin D. Virus was detected in patient lymphocytes and it was shown that a singular nuclear focus also characterizes cells infected in vivo. The cell line 8E5/LAV containing one defective integrated provirus revealed a similar focus of nuclear RNA, and the single integrated HIV genome was unequivocally visualized on a D-group chromosome. This demonstrates an extremely sensitive single-cell assay for the presence of a single site of HIV transcription in vitro and in vivo and suggests that it derives from one (or very few) viral genomes per cell. In contrast, productive Epstein-Barr virus infection exhibited many foci of nuclear RNA per cell.</p>
dc.identifier.submissionpathoapubs/1830
dc.contributor.departmentDepartment of Pediatrics
dc.contributor.departmentDepartment of Cell Biology
dc.source.pages5420-4


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