Cross-reaction of hnRNP-proteins of HeLa cells with nuclear proteins of Drosophila melanogaster demonstrated by a monoclonal antibody*1

Hügle, Barbara; Guldner, H. H.; Bautz, Friedlinde A.; Alonso, Ángel

doi:10.1016/0014-4827(82)90416-5

Cited by 13 publications

(3 citation statements)

References 10 publications

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“…Many of these were found by immunofluorescence to decorate dencondensed regions of polytene chromosomes. Some of the respective antigens were associated with pulse-labeled RNA (Risau et al, 1982;Hugle et al, 1982).…”

Section: Introductionmentioning

confidence: 99%

“…Such studies should also yield information on the question of whether there are different RNPs associated with different species of heterogeneous nuclear RNA (hnRNA). Therefore, we have tried to localize, by indirect immunofluorescence using monoclonal antibodies, a set of NHCPs which were found to be associated with labeled RNA (Hugle et al, 1982;Risau et al, in preparation) on polytene chromosomes of D. melanogaster.…”

Section: Introductionmentioning

confidence: 99%

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Differential distribution of RNA polymerase B and nonhistone chromosomal proteins in polytene chromosomes of Drosophila melanogaster.

Kabisch

Bautz

1983

The EMBO Journal

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The distribution patterns of nonhistone chromosomal proteins (NHCP) associated with pulse‐labeled RNA were determined by indirect immunofluorescence on salivary gland chromosomes of Drosophila melanogaster using monoclonal antibodies. By staining for two different antigens simultaneously, using antibodies tagged with different fluorescent probes, it became possible to position RNA‐associated antigens as well as RNA polymerase B in relation to each other. Three separate staining patterns could be observed with anti‐NHCP antibodies, none of which showed a pattern which was identical with that of RNA polymerase B. Furthermore, no correlation with the synthesis of the primary trancript, as monitored by the RNA polymerase B content of chromosomal sites, could be found by following the fluorescence patterns during inactivation of intermolt puffs or activation of early ecdysone‐induced puffs. Finally, no strict correlation was observed between puffing activity and the accumulation of a certain antigen in these selected chromosomal sites.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Differential distribution of RNA polymerase B and nonhistone chromosomal proteins in polytene chromosomes of Drosophila melanogaster.

Kabisch

Bautz

1983

The EMBO Journal

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“…1), as far as could be discerned, no cytoplasmic fluorescence was evident. Nor is the nuclear staining so diffuse as found for heterogeneous nuclear RNA-ribonucleoprotein antigens (26). The staining patterns are always highly reproducible within and between experiments.…”

Section: Cellular Localization and Distribution Pattern Of The Antigens By Indirect Immunofluorescencementioning

confidence: 86%

Monoclonal antibodies specific for tight-binding human chromatin antigens reveal structural rearrangements within the nucleus during the cell cycle.

Bhorjee¹,

Barclay²,

Wedrychowski³

et al. 1983

The Journal of Cell Biology

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The class of nonhistone chromosomal proteins that remains bound to DNA in chromatin in the presence of 2.5 M NaCl-5 M urea has proven refractile to biochemical analysis. In order to study its role in chromatin organization, we have produced monoclonal antibodies that are specific for the HeLa DNA-protein complex that remains after extraction of chromatin with high salt and urea. The antibody-producing clones were identified with an ELISA assay. Of the six clones selected, five were stabilized by limiting dilution. All clones are IgG producers. None cross-react significantly with native DNA, core histones, or the highmobility group nonhistone proteins. All antibodies are specific for nuclear or juxtanuclear antigens. Indirect immunofluorescence shows that three antibodies, which are nonidentical, stain three different nuclear networks. Available evidence indicates that two of these networks are the nuclear matrix. A fourth antibody reveals structures reminiscent of chromocenters. A fifth antibody, AhNA-I, binds to interphase HeLa chromatin and specifically decorates metaphase chromosomes. AhNA-I similarly recognizes rat chromosomes. Each of these monoclonal antibodies also reveals a changing pattern of nuclear staining as cells progress through the cell cycle. Presumably, this reflects the rearrangement of the cognate antigens.The eucaryotic chromosome contains at least twice as much protein by mass as DNA and both are complexed in a structure defined as chromatin. Treatment of either purifed nuclei or chromatin by high salt (2.0 M NaC1) removes almost all histones and most nonhistone proteins (1-3). The proteins remaining bound to DNA in chromatin after high ionic strength extraction (2 M NaCI or 2 M NaC1-5M urea) are nonhistones, and they constitute 5-8% of the total chromatin protein. These are termed residual or tight-binding proteins (4-6). The tight-binding, nonhistone chromatin proteins (TBP) from animal cell nuclei are electrophoretically complex (~200 species, Bhorjee, J. S. and L. Kite, unpublished data), and are distributed nonrandomly along the DNA molecule (reference 5, reviewed in reference 6).Two functions for these proteins have been suggested, although no direct evidence exists. Based on the DNA sequencespecific association of certain nonhistone proteins with chromatin DNA, a role in specific gene expression for these proteins has been proposed (7-13). It has also been demonstrated that some TBPs bind to androgen-receptor (14) and progesterone-receptor (15) complexes. Thus, at least some gene regulatory molecules in eukaryotes may reside in this group of chromosomal proteins. A second function may be structural. The structural integrity of the nuclear matrix (4, 16) and the chromosome scaffold (17) seems to rely on the presence of such high-ionic strength (2 M NaCI), nonextractable nuclear nonhistone proteins.Closer examination of these proposals by biochemical analysis of individual tight-binding nonhistone proteins has not been possible for two reasoaas: (a) each is present in small am...

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Differential distribution of nonhistone proteins from polytene chromosomes of Drosophila melanogaster after heat shock

Dangli

Bautz

1983

Chromosoma

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Nine monoclonal antibodies directed against chromosomal proteins of D. melanogaster were used to study, by indirect immunofluorescence, the distribution of their respective antigens on polytene chromosomes following heat shock. This treatment is known to induce a specific set of transcriptionally active puffs with concomitant reduction of transcriptional activity in previously active loci. Our studies revealed wide differences in the distribution of the individual chromosomal proteins under heat shock conditions with regard to pattern and rate of both elimination from the inactivated loci and accumulation in the activated loci.

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Cross-reaction of hnRNP-proteins of HeLa cells with nuclear proteins of Drosophila melanogaster demonstrated by a monoclonal antibody*1

Cited by 13 publications

References 10 publications

Differential distribution of RNA polymerase B and nonhistone chromosomal proteins in polytene chromosomes of Drosophila melanogaster.

Differential distribution of RNA polymerase B and nonhistone chromosomal proteins in polytene chromosomes of Drosophila melanogaster.

Monoclonal antibodies specific for tight-binding human chromatin antigens reveal structural rearrangements within the nucleus during the cell cycle.

Differential distribution of nonhistone proteins from polytene chromosomes of Drosophila melanogaster after heat shock

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