HeLa cells and HeLa cells expressing the HIV-1 regulatory protein Rev were immunostained for Rev and pre-mRNA processing factors and examined histographically by confocal laser scanning microscopy. Following short pulse-labelling with bromouridine tri-phosphate nascent RNA gave a granular nucleoplasmic staining increasing somewhat towards the periphery as did also the heterogeneous ribonucleoproteins (hnRNPs) A1 and particularly C1/C2, a distribution pattern which has not been described. The sm-antigen of the small ribonucleoprotein particle (snRNP) proteins U1, U2, U4/U6 and U5 stained the nucleoplasm diffusely in addition to speckles which co-localised with speckles of the non-snRNP splicing factor SC-35. Brominated RNA and the hnRNPs A1 and C1/C2 were to varying degrees excluded from the speckles. Rev concentrated in the nucleolus and often as a perinucleolar ring/zone. Rev also stained the nucleoplasm and cytoplasm without co-localising with the above-mentioned proteins or brominated RNA and was not enriched or excluded in SC-35 speckles. The nucleolar proteins B23 and C23, like Rev, gave primarily a perinucleolar ring and stained the nucleoplasm but did not otherwise co-localise with Rev or with nuclear proteins. Histographic recording of immunofluorescence images proved to be a valuable tool in the study of localisation of HIV-1 Rev and cellular components and of possible co-localisations. A parallel comparison of the subcellular patterns of pre-mRNA processing factors versus major nucleolar antigens is new and suggests that the factors are not strictly separated in the nucleoplasm.
http://linkedlifedata.com/resource/pubmed/chemical/Bromodeoxyuridine, http://linkedlifedata.com/resource/pubmed/chemical/Gene Products, rev, http://linkedlifedata.com/resource/pubmed/chemical/Nuclear Proteins, http://linkedlifedata.com/resource/pubmed/chemical/RNA, Messenger, http://linkedlifedata.com/resource/pubmed/chemical/RNA, Viral, http://linkedlifedata.com/resource/pubmed/chemical/rev Gene Products, Human...
pubmed-meshheading:9541613-Animals, pubmed-meshheading:9541613-Bromodeoxyuridine, pubmed-meshheading:9541613-Fluorescent Antibody Technique, pubmed-meshheading:9541613-Gene Products, rev, pubmed-meshheading:9541613-HIV-1, pubmed-meshheading:9541613-HeLa Cells, pubmed-meshheading:9541613-Humans, pubmed-meshheading:9541613-Immunohistochemistry, pubmed-meshheading:9541613-Mice, pubmed-meshheading:9541613-Nuclear Proteins, pubmed-meshheading:9541613-RNA, Messenger, pubmed-meshheading:9541613-RNA, Viral, pubmed-meshheading:9541613-Transfection, pubmed-meshheading:9541613-rev Gene Products, Human Immunodeficiency Virus
Histographic recording of human immunodeficiency virus type 1 (HIV-1) regulatory protein Rev and nuclear factors.
Department of Microbiology and Immunology, Gade Institute, University of Bergen, High Technology Center of Bergen, Norway.
Journal Article, Research Support, Non-U.S. Gov't