Identification of a Nuclear Ribonucleoprotein Particle Which Contains Incompletely Spliced HIV-1 RNAs

• Published in: Biochemical and biophysical research communications Vol. 268; no. 2; pp. 509 - 513
• Main Authors: Bøe, Stig Ove, Kalland, Karl-Henning
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 16-02-2000
• Subjects: 100S particles; AIDS/HIV; b-actin; glyceraldehyde-3-phosphate dehydrogenase; HIV-1 - genetics; HIV-1 - metabolism; Human immunodeficiency virus 1; Humans; Lymphocytes - virology; nuclear extracts; Rev protein; Ribonucleoproteins, Small Nuclear - chemistry; RNA Splicing - genetics; RNA, Viral - genetics; RNA, Viral - metabolism; sucrose gradient centrifugation; Tumor Cells, Cultured; nuclear extracts; 100S particles; sucrose gradient centrifugation
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1006/bbrc.2000.2177

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 β-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.