Drug-induced dispersal of transcribed rRNA genes and transcriptional products: Immunolocalization and silver staining of different nucleolar components in rat cells treated with 5,6-dichloro-β-D-ribofuranosylbenzimidazole

U. Scheer, B. Hugle, Rachel Hazan, K. M. Rose

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Abstract

Upon incubation of cultured rat cells with the adenosine analogue 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole (DRB), nucleoli reversibly dissociate into their substructures, disperse throughout the nuclear interior, and form nucleolar 'necklaces'. We have used this experimental system, which does not inhibit transcription of the rRNA genes, to study by immunocytochemistry the distribution of active rRNA genes and their transcriptional products during nucleolar dispersal and recovery to normal morphology. Antibodies to RNA polymerase I allow detection of template-engaged polymerase, and monoclonal antibodies to a ribosomal protein (S1) of the small ribosomal subunit permit localization of nucleolar preribosomal particles. The results show that, under the action of DRB transcribed rRNA, genes spread throughout the nucleoplasm and finally appear in the form of several rows, each containing several (up to 30) granules positive for RNA polymerase I and argyrophilic proteins. Nucleolar material containing preribosomal particles also appears in granular structures spread over the nucleoplasm but its distribution is distinct from that of rRNA gene-containing granules. We conclude that, although transcriptional units and preribosomal particles are both redistributed in response to DRB, these entities retain their individuality as functionally defined subunits. We further propose that each RNA polymerase-positive granular unit represents a single transcription unit and that each continuous array of granules ('string of nucleolar beads') reflects the linear distribution of rRNA genes along a nucleolar organizer region. Based on the total number of polymerase I-positive granules we estimate that a minimum of 60 rRNA genes are active during interphase of DRB-treated rat cells.

Original languageEnglish (US)
Pages (from-to)672-679
Number of pages8
JournalJournal of Cell Biology
Volume99
Issue number2
StatePublished - 1984
Externally publishedYes

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Silver Staining
rRNA Genes
Pharmaceutical Preparations
RNA Polymerase I
Small Ribosome Subunits
Nucleolus Organizer Region
Interphase
DNA-Directed RNA Polymerases
Individuality
Adenosine
Cultured Cells
Immunohistochemistry
Monoclonal Antibodies
Antibodies
Proteins

ASJC Scopus subject areas

  • Cell Biology

Cite this

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title = "Drug-induced dispersal of transcribed rRNA genes and transcriptional products: Immunolocalization and silver staining of different nucleolar components in rat cells treated with 5,6-dichloro-β-D-ribofuranosylbenzimidazole",
abstract = "Upon incubation of cultured rat cells with the adenosine analogue 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole (DRB), nucleoli reversibly dissociate into their substructures, disperse throughout the nuclear interior, and form nucleolar 'necklaces'. We have used this experimental system, which does not inhibit transcription of the rRNA genes, to study by immunocytochemistry the distribution of active rRNA genes and their transcriptional products during nucleolar dispersal and recovery to normal morphology. Antibodies to RNA polymerase I allow detection of template-engaged polymerase, and monoclonal antibodies to a ribosomal protein (S1) of the small ribosomal subunit permit localization of nucleolar preribosomal particles. The results show that, under the action of DRB transcribed rRNA, genes spread throughout the nucleoplasm and finally appear in the form of several rows, each containing several (up to 30) granules positive for RNA polymerase I and argyrophilic proteins. Nucleolar material containing preribosomal particles also appears in granular structures spread over the nucleoplasm but its distribution is distinct from that of rRNA gene-containing granules. We conclude that, although transcriptional units and preribosomal particles are both redistributed in response to DRB, these entities retain their individuality as functionally defined subunits. We further propose that each RNA polymerase-positive granular unit represents a single transcription unit and that each continuous array of granules ('string of nucleolar beads') reflects the linear distribution of rRNA genes along a nucleolar organizer region. Based on the total number of polymerase I-positive granules we estimate that a minimum of 60 rRNA genes are active during interphase of DRB-treated rat cells.",
author = "U. Scheer and B. Hugle and Rachel Hazan and Rose, {K. M.}",
year = "1984",
language = "English (US)",
volume = "99",
pages = "672--679",
journal = "Journal of Cell Biology",
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TY - JOUR

T1 - Drug-induced dispersal of transcribed rRNA genes and transcriptional products

T2 - Immunolocalization and silver staining of different nucleolar components in rat cells treated with 5,6-dichloro-β-D-ribofuranosylbenzimidazole

AU - Scheer, U.

AU - Hugle, B.

AU - Hazan, Rachel

AU - Rose, K. M.

PY - 1984

Y1 - 1984

N2 - Upon incubation of cultured rat cells with the adenosine analogue 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole (DRB), nucleoli reversibly dissociate into their substructures, disperse throughout the nuclear interior, and form nucleolar 'necklaces'. We have used this experimental system, which does not inhibit transcription of the rRNA genes, to study by immunocytochemistry the distribution of active rRNA genes and their transcriptional products during nucleolar dispersal and recovery to normal morphology. Antibodies to RNA polymerase I allow detection of template-engaged polymerase, and monoclonal antibodies to a ribosomal protein (S1) of the small ribosomal subunit permit localization of nucleolar preribosomal particles. The results show that, under the action of DRB transcribed rRNA, genes spread throughout the nucleoplasm and finally appear in the form of several rows, each containing several (up to 30) granules positive for RNA polymerase I and argyrophilic proteins. Nucleolar material containing preribosomal particles also appears in granular structures spread over the nucleoplasm but its distribution is distinct from that of rRNA gene-containing granules. We conclude that, although transcriptional units and preribosomal particles are both redistributed in response to DRB, these entities retain their individuality as functionally defined subunits. We further propose that each RNA polymerase-positive granular unit represents a single transcription unit and that each continuous array of granules ('string of nucleolar beads') reflects the linear distribution of rRNA genes along a nucleolar organizer region. Based on the total number of polymerase I-positive granules we estimate that a minimum of 60 rRNA genes are active during interphase of DRB-treated rat cells.

AB - Upon incubation of cultured rat cells with the adenosine analogue 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole (DRB), nucleoli reversibly dissociate into their substructures, disperse throughout the nuclear interior, and form nucleolar 'necklaces'. We have used this experimental system, which does not inhibit transcription of the rRNA genes, to study by immunocytochemistry the distribution of active rRNA genes and their transcriptional products during nucleolar dispersal and recovery to normal morphology. Antibodies to RNA polymerase I allow detection of template-engaged polymerase, and monoclonal antibodies to a ribosomal protein (S1) of the small ribosomal subunit permit localization of nucleolar preribosomal particles. The results show that, under the action of DRB transcribed rRNA, genes spread throughout the nucleoplasm and finally appear in the form of several rows, each containing several (up to 30) granules positive for RNA polymerase I and argyrophilic proteins. Nucleolar material containing preribosomal particles also appears in granular structures spread over the nucleoplasm but its distribution is distinct from that of rRNA gene-containing granules. We conclude that, although transcriptional units and preribosomal particles are both redistributed in response to DRB, these entities retain their individuality as functionally defined subunits. We further propose that each RNA polymerase-positive granular unit represents a single transcription unit and that each continuous array of granules ('string of nucleolar beads') reflects the linear distribution of rRNA genes along a nucleolar organizer region. Based on the total number of polymerase I-positive granules we estimate that a minimum of 60 rRNA genes are active during interphase of DRB-treated rat cells.

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