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PMID: 6322105 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The in vivo and in vitro initiation site for transcription of the rRNA operon of Saccharomyces carlsbergensis.

Nucleic acids research ·Vol. 12 ·No. 3 ·1984-02-10 ·Pages 1377-90

Klootwijk J, Verbeet MP, Veldman GM, de Regt VC, van Heerikhuizen H, Bogerd J, Planta RJ

Abstract

We have performed a detailed analysis of the transcription initiation of the rRNA operon in the yeast Saccharomyces carlsbergensis. Electron microscopic analysis of R-looped pre-rRNA molecules together with a very sensitive S1-nuclease mapping showed the use of only a single transcription start at about 700 bp upstream of the 17S rRNA gene and not of the minor start sites proposed for the very closely related species S. cerevisiae by others [Bayev et al. (5), Swanson and Holland (6)]. The sequence of 730 bp of the initiating region is presented. In vitro transcription in concentrated lysates of yeast spheroplasts in the presence of (gamma-SH)ATP or (gamma-SH)GTP, followed by purification of the in vitro initiated RNA via Hg-agarose, revealed that on the endogenous template exactly the same site is used for transcription initiation as in vivo.

MeSH Terms

Base Sequence DNA/genetics DNA Restriction Enzymes DNA, Ribosomal Endonucleases Genes, Fungal Kinetics Nucleic Acid Hybridization Operon RNA, Ribosomal/genetics Saccharomyces/genetics Single-Strand Specific DNA and RNA Endonucleases Spheroplasts/metabolism Templates, Genetic Transcription, Genetic

Chemicals

DNA, Ribosomal RNA, Ribosomal DNA Endonucleases DNA Restriction Enzymes Single-Strand Specific DNA and RNA Endonucleases

Authors & Affiliations

7 authors, click to expand affiliations / ORCID

Klootwijk J

Verbeet M P

Veldman G M

de Regt V C

van Heerikhuizen H

Bogerd J

Planta R J

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21 references, click to expand

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Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/12.3.1377

Published

1984-02-10

Pages

1377-90

Language

English

Region

England

NLM ID

0411011

PMCID

PMC318583

Subset

Databases

GENBANK

X00380

External Links

PubMed source DOI: 10.1093/nar/12.3.1377 PMC Full Text View journal details

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The in vivo and in vitro initiation site for transcription of the rRNA operon of Saccharomyces carlsbergensis.

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