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121 results on '"Puvion-Dutilleul, Francine"'

11. Anchorage of Adenoviral RNAs to Clusters of Interchromatin Granules

Author

Besse, Sylvie and Puvion-Dutilleul, Francine

Subjects
viruses, Adenoviruses, Human, RNA, Small Nuclear, Spliceosomes, Humans, RNA, Viral, RNA, Messenger, DNA Probes, Article, Chromatin, In Situ Hybridization, HeLa Cells
Abstract

Previous in situ hybridization experiments have revealed that clusters of interchromatin granules in adenovirus type 5 infected HeLa cells contain not only spliceosome components but also significant amounts of viral RNA and poly(A)+ RNA molecules whereas nonpolyadenylated viral RNA molecules are present within the still enigmatic viral compact rings. To determine the levels of association of the viral RNA molecules with cellular clusters of interchromatin granules and viral compact rings, we investigated the effects of a cell extraction technique on these structures. The spreading apart of the nucleoproteins by exposure of infected cells to a detergent-containing hypotonic solution, which exclusively preserves structurally linked components, resulted in the persistence within the clusters of interchromatin granules of U1 snRNA, U2 snRNA, viral RNA, and poly(A)+ RNA. These data clearly reveal that, in addition to the well-known strong binding of spliceosome components within the clusters of interchromatin granules, there also is an anchorage of viral RNA and messenger RNA molecules to these structures, which suggests functional relationships. Taken together, the data indicate that the clusters of interchromatin granules might be the sites of accumulation and retention of those cell and viral messenger RNA molecules that are transiently stored in the nucleus before their degradation or their transport to the cytoplasm. In addition, the firm binding of nonpolyadenylated viral RNA to the viral compact rings suggests a role for these structures in the transient storage of the nonused portions of the viral primary late transcripts.

Published
2018

22. Isolation and Characterization of an Equine Foamy Virus

Author

Tobaly-Tapiero, Joelle, primary, Bittoun, Patricia, additional, Neves, Manuel, additional, Guillemin, Marie-Claude, additional, Lecellier, Charles-Henri, additional, Puvion-Dutilleul, Francine, additional, Gicquel, Bernard, additional, Zientara, Stephan, additional, Giron, Marie-Louise, additional, de Thé, Hugues, additional, and Saïb, Ali, additional

Published
2000
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46. LOCALIZATION OF U1, U2 snRNAs AND POLY(A) RNA BY IN SITU HYBRIDIZATION AT THE ELECTRON MICROSCOPE LEVEL. CHARACTERIZATON OF A NOVEL NUCLEAR DOMAIN CONTAINING U1 but not U2 snRNA AND PRESENCE OF POLY(A) RNA IN INTERCHROMATIN GRANULES

Author

Puvion-Dutilleul Francine, Puvion Edmond, Bachellerie Jean-Pierre, Harper Francis, and Visa Neus

Subjects
law, Biophysics, Poly-A RNA, Cell Biology, General Medicine, In situ hybridization, Electron microscope, Biology, Molecular biology, Small nuclear RNA, law.invention, Domain (software engineering)
Published
1993

48. Distribution of Low-Molecular-Weight RNAs in Avian Erythroblast Nuclear Ribonucleoprotein Complexes Associated with Pre-messenger RNA.

Author

Maxwell, E. Stuart, Maundrell, Kinsey, Puvion-Dutilleul, Francine, and Scherrer, Klaus

Subjects
RNA, NUCLEOPROTEINS, LYSOSOMES, NUCLEOTIDES, PROTEINS, BIOCHEMISTRY
Abstract

Duck erythroblast nuclei, isolated under conditions designed to preserve lysosome integrity, were extracted to yield a bulk population of nuclear ribonucleoprotein (nRNP) particles which. when fractionated on sucrose gradients, was resolved into two classes of RNA-protein complexes containing low-molecular-weight RNA. The classical pre-messenger ribonucleoprotein complex (pre-mRNP), which had been previously characterized in this laboratory and was shown to contain rapidly-labeled pre-messenger RNA (pre-mRNA) sequences, was observed to contain approximately eight discrete species of nuclear RNA between 90 and 230 nucleotides in length (major species of Mr 80000 and 55000, and minor species of Mr 61000, 58000, 51000, 38000, 36000 and 32000). The size and relative distribution of these RNA sequences indicated that they were the small nuclear RNA species previously described in the literature. In addition, two smaller ribonucleoprotein complexes with sedimentation coefficients of 17 S and 12 S were characterized and shown to be composed of characteristic, although not mutually exclusive, sets of low-Mr RNA species. Comparison of these RNA species in the two smaller nRNP complexes with those found in the 40-S pre-mRNP particles demonstrated that all but three of the major low-Mr RNA bands found in the small nRNP were found in the pre-mRNP. The additional RNA species unique to the small nRNP were major ones of Mr 150000 and 110000, minor ones of Mr 48000, 34000 and 27000 and three major bands of Mr approximately 25000 as well. Further comparison revealed that while each particle was composed of a characteristic subset of polypeptides, several common proteins were observed in both small nRNP and pre-mRNP. The common RNA and polypeptide components observed in these two classes of nuclear ribonucleoprotein suggested that these small nRNPs may represent either integral parts of intact physiological pre-mRNP or nuclear structures which are closely associated with pre-mRNP in vivo. No difference was observed in either the RNA or protein composition of small nRNP or pre-mRNP when prepared in the presence of the protease inhibitor phenylmethylsulfonyl fluoride. indicating that endogenous protease activity was not a problem in the preparation of nRNP from duck erythroblasts. CsCl gradient analysis of the two fractionated small nRNPs revealed buoyant densities of 1.39 and 1.40 g/ml for the IT-S and 12-S particles respectively. From these densities, an approximate RNA:protein ratio of 1:4, similar to that previously determined for the 40-S pre-mRNP complex, was estimated. Isoelectric focusing of the small nRNP on a pH gradient between pH 4 and pH 6 and electron microscopy studios reaffirmed the particulate nature of these smaller ribonucleoprotein complexes. In addition, ultrastructural analysis of small nRNP particles indicated an average diameter of 15 nm, compared with an average diameter of 35–40 nm for the bulk of the 40-S pre-mRNP particles. The poly(A)-protein complex of pre-RNA, sedimenting at a position of 15 S on the sucrose gradients, was bound to oligo(dT)-cellulose and subsequently eluted from the column in buffer containing 50% formamide. Six major proteins were associated with this poly(A) sequence, including the 73000-Mr poly(A)-binding protein typically seen associated with the poly(A) sequence of mRNA and pre-mRNA. In contrast, neither the 17-S nor the 12-S nRNP was bound to the oligo(dT)cellulose column indicating that these nRNPs did not contain poly(A) sequences and were, therefore, nRNP complexes distinctly different from the previously described poly(A)-protein complex of pre-mRNA. [ABSTRACT FROM AUTHOR]

Published
1981
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49. p80-coilin: a component of coiled bodies and interchromatin granule-associated zones

Author

Puvion-Dutilleul, Francine, Besse, Sylvie, Chan, Edward K. L., Tan, Eng M., and Puvion, Edmond

Abstract

We investigated at the electron microscope level the fate of the three intranuclear structures known to accumulate snRNPs, and which correspond to the punctuate immuno-fluorescent staining pattern (the coiled bodies, the clusters of interchromatin granules and the interchromatin granule-associated zones) after exposure to either a low salt medium which induces a loosening and partial spreading of nucleoprotein fibers or a high ionic strength salt medium and subsequent DNase I digestion, in order to obtain DNA-depleted nuclear matrices. The loosened clusters of inter-chromatin granules and the coiled bodies could no longer be distinguished from surrounding nucleoprotein fibers solely by their structure, but constituents of the clusters of interchromatin granules could be detected by in situ hybridization with both U1 and U2 DNA probes, and con-stituents of the coiled bodies were detectable mainly with the U2 DNA probe. The interchromatin granule-associated zones, the electron-opacity and compactness of which were preserved despite the loosening treatment, remained labeled with the U1 DNA probe only. In DNA-depleted nuclear matrices, the snRNA content of the coiled bodies, the clusters of interchromatin granules and their associated zones, which were all easily recognizable within the residual nuclear ribonucleoprotein network, was unmodi-fied. The data indicate, therefore, that the loosening procedure as well as the high salt extraction procedure preserve the snRNA content of all three spliceosome component-accumulation sites and reveal that interchro-matin granule-associated zones are elements of the nuclear matrix. The p80-coilin content of coiled bodies was also preserved whatever the salt treatment used. An intriguing new finding is the detection of abundant p80-coilin within the interchromatin granule-associated zones, both before and after either low or high salt treatment of cells. Therefore, p80-coilin is an integral constituent of the inter-chromatin granule-associated zones.

Published
1995
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50. Rearrangements of intranuclear structures involved in RNA processing in response to adenovirus infection

Author

Puvion-Dutilleul, Francine, Bachellerie, Jean-Pierre, Visa, Neus, and Puvion, Edmond

Abstract

We have studied in HeLa cells at the electron microscope level the response to adenovirus infection of the RNA pro- cessing machinery. Components of the spliceosomes were localized by in situ hybridization with biotinylated U1 and U2 DNA probes and by immunolabeling with Y12 anti-Sm monoclonal antibody, whereas poly(A)+ RNAs were localized by specific binding of biotinylated poly(dT) probe. At early stages of nuclear transformation, the distribution of small nuclear RNPs was similar to that previously described in non-infected nuclei (Visa, N., Puvion-Dutilleul, F., Bachellerie, J. P. and Puvion, E., Eur. J. Cell Biol. 60, 308-321, 1993; Visa, N., Puvion-Dutilleul, F., Harper, F., Bachellerie, J. P. and Puvion, E., Exp. Cell Res. 208, 19-34, 1993). As the infection progresses, the large virus-induced inclusion body consists of a central storage site of func- tionally inactive viral genomes surrounded by a peripheral shell formed by clusters of interchromatin granules, compact rings and a fibrillogranular network in which are embedded the viral single-stranded DNA accumulation sites. Spliceosome components and poly(A)+ RNAs were then exclusively detected over the clusters of interchro- matin granules and the fibrillogranular network whereas the viral single-stranded DNA accumulation sites and compact rings remained unlabeled, thus appearing to not be directly involved in splicing. Our data, therefore, suggest that the fibrillogranular network, in addition to being the site of viral transcription, is also a major site of viral RNA splicing. Like the clusters of interchromatin granules, which had been already involved in spliceosome assembly, they could also have a role in the sorting of viral spliced polyadenylated mRNAs before export to the cytoplasm. The compact rings, which contain non- polyadenylated viral RNA, might accumulate the non-used portions of the viral transcripts resulting from differential poly(A)+ site selection.

Published
1994
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