Identification of a nuclear ribonucleoprotein particle which contains incompletely spliced HIV-1 RNAs.
Unspliced and partially spliced HIV RNAs are transported to the cytoplasm by the HIV encoded Rev protein. In the present study, a ribonucleoprotein complex which contains such incompletely spliced HIV RNA is identified. Soluble nuclear extracts were prepared from the lymphocyte cell line H9/IIIB that constitutively produces HIV-1 from a stably integrated provirus. Sucrose gradient centrifugation of the extracts and subsequent analysis of the gradient fractions by a ribonuclease protection assay revealed a population of incompletely spliced HIV-1 RNAs which accumulates in the 100S region of the gradient. Similar analysis of cellular mRNAs including beta-actin and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) revealed that these RNA molecules also exhibit characteristic sedimentation profiles in sucrose gradients. This study suggests that nuclear ribonucleoprotein particles containing incompletely spliced HIV-1 RNAs are amenable for biochemical characterisation.
Bøe
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Center for Research in Virology, Department of Microbiology and Immunology, The Gade Institute, University of Bergen, Bergen High Technology Center, Bergen, N-5020, Norway. Stig.boe@vir.uib.no
Kalland
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D015497:HIV-1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008214:Lymphocytes / Q000821:virology; D012326:RNA Splicing / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D017411:Ribonucleoproteins, Small Nuclear / Q000737:chemistry*; D014407:Tumor Cells, Cultured
D012367:RNA, Viral; D017411:Ribonucleoproteins, Small Nuclear