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

J. Klootwijk; M.P. Verbeet; G.M. Veldman; V.C. de Regt; H. van Heerikhuizen; J. Bogerd; R.J. Planta

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

J. Klootwijk, M.P. Verbeet, G.M. Veldman, V.C. de Regt, H. van Heerikhuizen, J. Bogerd, R.J. Planta

Research output: Contribution to journal › Article › Academic › peer-review

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.

Original language	English
Pages (from-to)	1377-1390
Number of pages	14
Journal	Nucleic Acids Research
Volume	12
Issue number	3
Publication status	Published - 1984

Keywords

DNA
endonuclease
nuclease S1
restriction endonuclease
ribosome DNA
ribosome RNA
article
DNA template
fungal gene
genetic transcription
genetics
kinetics
metabolism
nucleic acid hybridization
nucleotide sequence
operon
Saccharomyces
spheroplast

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@article{10e56d5399be446d85af07772bed5c64,

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

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.",

keywords = "DNA, endonuclease, nuclease S1, restriction endonuclease, ribosome DNA, ribosome RNA, article, DNA template, fungal gene, genetic transcription, genetics, kinetics, metabolism, nucleic acid hybridization, nucleotide sequence, operon, Saccharomyces, spheroplast",

author = "J. Klootwijk and M.P. Verbeet and G.M. Veldman and {de Regt}, V.C. and {van Heerikhuizen}, H. and J. Bogerd and R.J. Planta",

year = "1984",

language = "English",

volume = "12",

pages = "1377--1390",

journal = "Nucleic Acids Research",

issn = "0305-1048",

publisher = "Oxford University Press",

number = "3",

}

TY - JOUR

T1 - The in vivo and in vitro initiation site for transcription of the rRNA operon of Saccharomyces carlsbergensis.

AU - Klootwijk, J.

AU - Verbeet, M.P.

AU - Veldman, G.M.

AU - de Regt, V.C.

AU - van Heerikhuizen, H.

AU - Bogerd, J.

AU - Planta, R.J.

PY - 1984

Y1 - 1984

N2 - 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.

AB - 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.

KW - DNA

KW - endonuclease

KW - nuclease S1

KW - restriction endonuclease

KW - ribosome DNA

KW - ribosome RNA

KW - article

KW - DNA template

KW - fungal gene

KW - genetic transcription

KW - genetics

KW - kinetics

KW - metabolism

KW - nucleic acid hybridization

KW - nucleotide sequence

KW - operon

KW - Saccharomyces

KW - spheroplast

M3 - Article

SN - 0305-1048

VL - 12

SP - 1377

EP - 1390

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 3

ER -

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

Abstract

Keywords

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