Computer Analysis of the Distribution of Nuclear Antigens: Studies on the Spatial and Functional Organization of the Interphase Nucleus

Ringertz, Nils R.; Hadlaczky, Gyula; Hallman, Heikki; Nyman, Ulf; Pettersson, Ingvar; Sharp, Gordon C.

doi:10.1242/jcs.1986.supplement_4.2

Cited by 33 publications

(16 citation statements)

References 63 publications

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“…All 5 non-nucleolar snRNAs have been identified in lymphocytes [43], and it has been shown that the Sm antigen is already present in relatively high amounts in resting lymphocytes [8]. Other studies using immunolabelling of protein [51,60] and RNA [50] components of snRNPs suggest that all snRNPs are colocalized throughout the mitotic cell cycle, as monitored by fluorescence microscopy, and also indicate that the snRNAs remain associated in complexes with the same proteins during the cell cycle. These data, together with the consistent detectioa of punctate peripheral labelling with PII but not with anti-snRNP, suggest that the PI1 antigen is not part of the snRNP particles and is therefore unlikely to be directly involved in splicing.…”

Section: Discussionmentioning

confidence: 98%

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Modulation of lymphocyte nuclear matrix organization in vivo by 5,6-dichloro-1-β-D-ribofuranosyl benzimidazole: an autoradiographic and immunofluorescence study

Chaly

1988

Biology of the Cell

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Assembly of active nuclei in lymphocytes stimulated by mitogen is paralleled by the elaboration of a structurally and biochemically complex nuclear matrix (NM). To examine the dynamics of individual NM polypeptide components during blastogenesis, we have applied immunofluorescence labelling with anti-NM antibodies to concanavalin A-stimulated mouse splenocytes. Whereas peripherin and PI2 antigens did not reorganize during stimulation, labelling of PI1 and small nuclear ribonucleoprotein (snRNP) antigens increased markedly in intensity and redistributed in concert with the previously reported NM restructuring. Double-labelling showed, furthermore, that snRNPs and the internal staining component of PI1 were largely co-localized. As an approach to studying the role of RNA and RNA synthesis in NM organization, we have further examined the effects of the inhibitor of RNA synthesis, 5,6-dichloro-1-beta-D-ribofuranosyl benzimidazole (DRB), on NM antigen distribution. The rapid inhibition of 3H-uridine incorporation by DRB was accompanied by coordinate aggregation of snRNPs and of the internal PI1 component into large, brightly stained patches. Both 3H-uridine incorporation levels and antigen localization were readily reversed upon removal of DRB. We conclude that NM antigens behave independently during nuclear and NM assembly and that NM organization, as reflected by NM antigen distribution, is modulated by con A- and DRB-induced alterations in RNA synthesis. We propose, furthermore, that the PI1 antigen plays a role in RNA metabolism, and is possibly involved in RNA transport to the nuclear periphery.

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Section: Discussionmentioning

confidence: 98%

“…The latter detects the Sm antigen common to all classes of snRNP particles [39,46], and results in staining patterns indistinguishable from those produced by anti-U1 snRNP (Fig. 2) [51]. In both type II (Figs.…”

Section: Nm Antigen Distribution During Mitogenic Stimulationmentioning

confidence: 92%

Modulation of lymphocyte nuclear matrix organization in vivo by 5,6-dichloro-1-β-D-ribofuranosyl benzimidazole: an autoradiographic and immunofluorescence study

Chaly

1988

Biology of the Cell

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“…Second, the 3D strueture and arrangement of chromatin may also be altered by pathological influences [28, [131][132][133]. Third, recent evidence has suggested the nonrandom distribution of nonchromosomal nuclear struetures, such as RNP particles [134]. In addition, fluorescence in situ hybridization experiments have demonstrated that RNA transcripts are accumulated and transported in a topologically highly ordered manner [135,136].…”

Section: Nuclear Topographymentioning

confidence: 99%

Analysis of genes and chromosomes by nonisotopic in situ hybridization

Lichter

Boyle

Cremer

et al. 1991

Genetic Analysis: Biomolecular Engineering

224

102

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“…These data in conjunction with those in the accompanying manuscript (Yang et al, 1992) lead to the hypothesis that an internal coiled-coil filamentous system may be a general structural component of the eukaryotic nucleus . (Ringertz et al, 1986) or clustered near the nucleolus (Bartholdi, 1991) .…”

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confidence: 99%

The NUF1 gene encodes an essential coiled-coil related protein that is a potential component of the yeast nucleoskeleton.

Mirzayan

Copeland

Snyder

1992

The Journal of Cell Biology

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Abstract. In an attempt to identify structural components of the yeast nucleus, subcellular fractions of yeast nuclei were prepared and used as immunogens to generate complex polyclonal antibodies . One such serum was used to screen a yeast genomic Xgtll expression library. A clone encoding a gene called NUF7 (for nuclear filament-related) was identified and extensively characterized . Antibodies to NUFI fusion proteins were generated, and affinity-purified antibodies were used for immunoblot analysis and indirect immunofluorescence localization . The NUF1 protein is 110 kD in molecular mass and localizes to the yeast nucleus in small granular patches. Intranuclear staining is present in cells at all stages of the cell cycle . The NUF1 protein of yeast is tightly associated with the nucleus ; it was not removed by extraction of nuclei with nonionic detergent or salt, or treatment with W 1THIN the cell nucleus, a variety of essential processes such as DNA replication, RNA transcription, RNA splicing, and tRNA Production occur (Newport and Forbes, 1987) . Recent evidence indicates that chromosomes and other components within the nucleus are often organized into morphological domains . The nucleolus, nuclear membrane, and splicing centers (or snurposomes) are sites of well-known nuclear functions (Carmo-Fonseca et al., 1991;Spector, 1990). Less well-characterized nuclear domains have also been described in mammalian cells. These include sites of more active gene transcription (defined by the presence of RNA-binding proteins in spread chromosomes [Alberts et al ., 1977;Igo-Kemenes et al., 1982]), nuclear "dots" [Ascoli and Maul, 1991; Xie, K., E. Lambie and M . Snyder, manuscript submitted for publication]), and nuclear bodies [Chaley et al., 1983 ;Vagner-Capodano et al ., 1982]) . In several species, chromosomes are also nonrandomly organized within the nucleus . For example, in Drosophila, polytene chromosomes of salivary glands have a specific configuration (Ellison and Howard, 1981 : Hochstrasser et al ., 1986;Saumweber, 1987), and in mammalian cells, centromeres can be paired, grouped at one end ofthe nucleus RNAse and DNAse . Sequence analysis of the NUR gene predicts a protein 945 amino acids in length that contains three domains: a large 627 residue central domain predicted to form a coiled-coil structure flanked by nonhelical amino-terminal and carboxyterminal regions . Disruption of the NUM gene indicates that it is necessary for yeast cell growth . These results indicate that NUF1 encodes an essential coiledcoil protein within the yeast nucleus ; we speculate that NUF1 is a component of the yeast nucleoskeleton . In addition, immunofluorescence results indicate that mammalian cells contain a NUFl-related nuclear protein. These data in conjunction with those in the accompanying manuscript (Yang et al ., 1992) lead to the hypothesis that an internal coiled-coil filamentous system may be a general structural component of the eukaryotic nucleus .( Ringertz et al., 1986) or clustered near the nucleolus (Barth...

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Computer Analysis of the Distribution of Nuclear Antigens: Studies on the Spatial and Functional Organization of the Interphase Nucleus

Cited by 33 publications

References 63 publications

Modulation of lymphocyte nuclear matrix organization in vivo by 5,6-dichloro-1-β-D-ribofuranosyl benzimidazole: an autoradiographic and immunofluorescence study

Modulation of lymphocyte nuclear matrix organization in vivo by 5,6-dichloro-1-β-D-ribofuranosyl benzimidazole: an autoradiographic and immunofluorescence study

Analysis of genes and chromosomes by nonisotopic in situ hybridization

The NUF1 gene encodes an essential coiled-coil related protein that is a potential component of the yeast nucleoskeleton.

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