Ultrastructural localization of nucleic acid sequences in Saccharomyces cerevisiae nucleoli

The putative nucleolus in Saccharomyces cerevisiae is visible in electron micrographs as a darkly stained, crescent-shaped structure associated with the nuclear envelope. The haploid yeast genome contains 100-200 tandem copies of a 9.1 kb ribosomal DNA (rDNA) repeat predicted to reside in this struc...

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Bibliographic Details
Published in:Chromosoma 1991-09, Vol.100 (8), p.519-523
Main Authors: Dvorkin, N, Clark, M.W, Hamkalo, B.A
Format: Article
Language:English
Subjects:
Base Sequence
Biological and medical sciences
cell nucleolus
Cell Nucleolus - chemistry
Cell Nucleolus - ultrastructure
cell ultrastructure
Cytogenetics
DNA probes
DNA, Fungal - analysis
DNA, Ribosomal - analysis
Fundamental and applied biological sciences. Psychology
Genetics of eukaryotes. Biological and molecular evolution
immunocytochemistry
Immunohistochemistry
in situ hybridization
Microscopy, Electron
Nucleic Acid Hybridization
repetitive sequences
ribosomal DNA
ribosomal RNA
RNA, Fungal - analysis
RNA, Ribosomal - analysis
Saccharomyces cerevisiae
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - ultrastructure
small nuclear RNA
Thallophyta, bryophyta
Vegetals and fungi
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Summary:The putative nucleolus in Saccharomyces cerevisiae is visible in electron micrographs as a darkly stained, crescent-shaped structure associated with the nuclear envelope. The haploid yeast genome contains 100-200 tandem copies of a 9.1 kb ribosomal DNA (rDNA) repeat predicted to reside in this structure. We combined in situ hybridization of non-isotopically labeled probes to isolated S. cerevisiae nuclei with immunogold detection to localize rDNA and rRNA precursor sequences in nuclei at the electron microscope (EM) level. Gold particles are restricted to defined regions of nuclei which appear more electron dense than the bulk of the nucleus and which generally exhibit the crescent shape typical of the structure thought to be the nucleolus. In addition, snR17, the yeast homolog of mammalian U3, a nucleolar-restricted small nuclear RNA (snRNA), was localized to the same electron dense region of the nucleus. These data, in conjunction with published immunofluorescent localizations of nucleolar-associated antigens, provide definitive proof that the dense crescent is the nucleolus. Finally, the technique described is applicable to probing nuclear organization in a genetically manipulable system.
ISSN:0009-5915
1432-0886
DOI:10.1007/BF00352202