In Situ Localization of RNA Structures

Puvion, Edmond; Moyne, Gilles

doi:10.1016/b978-0-12-147608-3.50008-2

Cited by 45 publications

(42 citation statements)

References 164 publications

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“…Interestingly, this study also recalled that the localization of replicating DNA at the periphery of condensed chromatin is similar to that observed for RNA synthesis, thereby giving further support to the long thought spatial colocalization between replication and transcription (Jaunin et al, 2000;Puvion and Moyne, 1981;Puvion and Puvion-Dutileul, 1996). Moreover, in mammalians cells, there is often a correlation between the transcriptional activity and the replication timing of a given gene (Pierron et al, 1999).…”

Section: Discussionsupporting

confidence: 78%

The relationship between BCL6 bodies and nuclear sites of normal and halogenated DNA and RNA synthesis

Puvion‐Dutilleul¹,

Souquere-Besse²,

Albagli-Curiel

2003

Microscopy Res & Technique

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BCL6 is a POZ/BTB and zinc finger transcription factor that self-interacts and accumulates into discrete nuclear "bodies" of unknown function. We recently reported that BCL6 bodies associate with bromodeoxyuridine (BrdU)-substituted DNA, suggesting their implication in replication. To examine this possibility, we examine here by electron and confocal microscopy the relation between BCL6 bodies and replication foci (RF) using incorporation of various halogenated nucleotides (BrdU, chlorodeoxyuridine, CldU, and iododeoxyuridine, IdU) or PCNA (proliferating cell nuclear antigen) staining. We show that BCL6 bodies are found associated with RF, as revealed by PCNA staining. However, such association is markedly prolonged upon BrdU or CldU incorporation, but less, or not at all, upon IdU incorporation. Pulse-chase and double-labeling experiments indicate that IdU-substituted DNA leaves BCL6 bodies after a few tenths of minutes while BrdUor CldU-substituted DNA stalls in their vicinity for several hours, thereby giving the characteristic "crowns" of DNA entirely surrounding BCL6 bodies. In all cases, however, the halogenated DNA ends up undergoing a movement from BCL6 bodies toward nucleoplasm and nuclear periphery to reach euchromatin and heterochromatin, respectively. We propose that replicating DNA is prone to be bound by BCL6, while BrdU/CldU incorporation increases this propensity possibly because these two events have synergistic effects on the structure and chromatinisation of the newly synthesized DNA. Finally, despite the known proximity between nuclear sites of transcription and replication, we show via several approaches that BCL6 bodies do not appear to be involved either in RNA synthesis or storage.

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Section: Discussionsupporting

confidence: 78%

The relationship between BCL6 bodies and nuclear sites of normal and halogenated DNA and RNA synthesis

Puvion‐Dutilleul¹,

Souquere-Besse²,

Albagli-Curiel

2003

Microscopy Res & Technique

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“…These structures, wh ich appeared irregularly scattered throughout the nucleoplasm, appeared as round ish aggregates of variable diameters (0.1-0.3 lAm) and revealed granular substructures of diameters 20-35 nm (Figures 11b and 11c). The morphology of these clustered particles closely resembles that of typical interchromatin granules (Swift, 1963;Monneron and Bernhard, 1969; for review see Puvion and Moyne, 1981). Obviously, these aggregates of 20-35 nm granules seen in the electron microscope correspond to the small fluorescent nucleoplasmic dots observed by light microscopic immunolocalization (compare Figures 9a and 10a' with Figures 11a-11d).…”

Section: Electron Microscopic Immunolocalization Of Ribocharinsupporting

confidence: 53%

Ribocharin: a nuclear Mr 40,000 protein specific to precursor particles of the large ribosomal subunit

Hügle

Scheer

Franke

1985

Cell

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SummaryUsing a monoclonal antibody (No-194) we have identified, in Xenopus laevis and other amphibia, an acidic protein of M, 40,000 (ribocharin) which is specifically associated with the granular component of the nucleolus and nucleoplasmic 65S particles. These particles contain the nuclear 28S rRNA and apparently represent the precursor to the large ribosomal subunit in nucleocytoplasmic transit. By immunoelectron microscopy ribocharin has been localized in the granular component of the nucleolus and in interchromatin granules. During mitosis ribocharin-containing particles are associated with surfaces of chromosomes and are recollected in the reconstituting nucleoli in late telophase. We suggest that ribocharin is a specific component of precursor particles of the large ribosomal subunit, which dissociates from the 65S particle before passage through the nuclear envelope, and is reutilized in ribosome biogenesis.

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“…Hsp7O accelerates changes in nucleolar morphology Nucleoli are very sensitive to heat shock; its effects have been documented in previous biochemical and ultrastructural studies (for reviews, see Simard et al, 1974;Puvion and Moyne, 1981). In electron micrographs, normal nucleoli are seen to consist of a fibrillar centre surrounded by a mass of fibrillar and granular RNPs.…”

Section: Resultsmentioning

confidence: 80%

Hsp70 accelerates the recovery of nucleolar morphology after heat shock.

Pelham¹

1984

The EMBO Journal

446

232

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The major heat-shock protein, hsp7O, is synthesized by cells of many organisms in response to stress. In the present study, Drosophila hsp7O was expressed from cloned genes in mouse L cells and monkey COS cells and detected by immunofluorescence using monoclonal antibodies. Hsp7O is found mostly but not exclusively in the nucleus of unstressed cells. For several hours after a short heat shock, however, it is strongly concentrated in nucleoli. Nucleoli are transiently damaged by such a heat shock: their morphology changes and assembly and export of ribosomes is blocked for several hours. This block can be visualized by addition of actinomycin D: under normal conditions pre-ribosomes are chased out of nucleoli, and the latter shrink dramatically, but no such shrinking is seen in heat-shocked cells. High levels of hsp7O can be produced in unstressed COS cells by transfecting them with an appropriate expression plasmid. Such cells show a more rapid recovery of nucleolar morphology following a heat shock than do untransfected cells. Furthermore, heat shock does not prevent shrinkage of their nucleoli in the presence of actinomycin, which indicates that ribosome export also recovers rapidly when pre-synthesized hsp7O is present. I suggest that an important function of hsp7O is to catalyze reassembly of damaged pre-ribosomes and other RNPs after heat shock.

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In Situ Localization of RNA Structures

Cited by 45 publications

References 164 publications

The relationship between BCL6 bodies and nuclear sites of normal and halogenated DNA and RNA synthesis

The relationship between BCL6 bodies and nuclear sites of normal and halogenated DNA and RNA synthesis

Ribocharin: a nuclear Mr 40,000 protein specific to precursor particles of the large ribosomal subunit

Hsp70 accelerates the recovery of nucleolar morphology after heat shock.

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