Open AccessThein vivoandin vitroinitiation site for transcription of the rRNA operon ofSaccharomyces carlsbergensis
Author(s) -
J. Klootwijk,
Martin Ph. Verbeet,
Geertruida M. Veldman,
V.C.H.F. de Regt,
Harm van Heerikhuizen,
Jan Bogerd,
Rudi J. Planta
Publication year - 1984
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/12.3.1377
Subject(s) - biology , operon , transcription (linguistics) , saccharomyces cerevisiae , saccharomyces , genetics , ribosomal rna , in vivo , in vitro , microbiology and biotechnology , gene , escherichia coli , linguistics , philosophy
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.