A cis-acting element present in multiple genes serves as a repressor protein binding site for the yeast CAR1 gene

R. M. Luche, R. Sumrada, Terrance Cooper

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

Induction of the arginase (CAR1) gene expression in Saccharomyces cerevisiae has previously been shown to require participation of a cis-dominantly regulated upstream repression sequence (URS). Deletion of this element results in high-level expression of the CAR1 gene without inducer. To determine the structure of the CAR1 URS element, we performed a saturation mutagenesis. Results of the mutagenic analysis indicated that the CAR1 URS was a 9-base-pair palindromic sequence, 5'-AGCCGCCGA-3'. A DNA fragment containing this sequence was shown to bind one or more proteins by a gel shift assay. DNA fragments containing point mutations that completely eliminated URS function were not effective competitors in this assay, whereas those which supported URS function were effective competitors. Sequences in the 5'-flanking regions of 14 other genes were found to be homologous to the CAR1 URS. These sequences were shown to support varying degrees of URS function in the expression vector assay, to bind protein as demonstrated by the gel shift assay, and to compete with a DNA fragment containing the CAR 1 URS for protein binding. These results indicate that the CAR1 URS element possesses the characteristics of a repressor binding site. These results indicate that the CAR1 URS element possesses the characteristics of a repressor binding site. Further, they are consistent with the suggestion that sites homologous to the CAR1 URS may be situated in the 5'-flanking regions of multiple unrelated yeast genes. The widespread occurrence of this element raises the possibility that it is the target site for one or more negatively acting general transcription factors.

Original languageEnglish (US)
Pages (from-to)3884-3895
Number of pages12
JournalMolecular and Cellular Biology
Volume10
Issue number8
DOIs
StatePublished - Jan 1 1990

Fingerprint

Repressor Proteins
Protein Binding
Yeasts
5' Flanking Region
Binding Sites
DNA
Gels
General Transcription Factors
Genes
Gene Expression
Arginase
Point Mutation
Mutagenesis
Base Pairing
Saccharomyces cerevisiae
Proteins

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

A cis-acting element present in multiple genes serves as a repressor protein binding site for the yeast CAR1 gene. / Luche, R. M.; Sumrada, R.; Cooper, Terrance.

In: Molecular and Cellular Biology, Vol. 10, No. 8, 01.01.1990, p. 3884-3895.

Research output: Contribution to journalArticle

@article{e71690dc97794b4493e14457e54df5e8,
title = "A cis-acting element present in multiple genes serves as a repressor protein binding site for the yeast CAR1 gene",
abstract = "Induction of the arginase (CAR1) gene expression in Saccharomyces cerevisiae has previously been shown to require participation of a cis-dominantly regulated upstream repression sequence (URS). Deletion of this element results in high-level expression of the CAR1 gene without inducer. To determine the structure of the CAR1 URS element, we performed a saturation mutagenesis. Results of the mutagenic analysis indicated that the CAR1 URS was a 9-base-pair palindromic sequence, 5'-AGCCGCCGA-3'. A DNA fragment containing this sequence was shown to bind one or more proteins by a gel shift assay. DNA fragments containing point mutations that completely eliminated URS function were not effective competitors in this assay, whereas those which supported URS function were effective competitors. Sequences in the 5'-flanking regions of 14 other genes were found to be homologous to the CAR1 URS. These sequences were shown to support varying degrees of URS function in the expression vector assay, to bind protein as demonstrated by the gel shift assay, and to compete with a DNA fragment containing the CAR 1 URS for protein binding. These results indicate that the CAR1 URS element possesses the characteristics of a repressor binding site. These results indicate that the CAR1 URS element possesses the characteristics of a repressor binding site. Further, they are consistent with the suggestion that sites homologous to the CAR1 URS may be situated in the 5'-flanking regions of multiple unrelated yeast genes. The widespread occurrence of this element raises the possibility that it is the target site for one or more negatively acting general transcription factors.",
author = "Luche, {R. M.} and R. Sumrada and Terrance Cooper",
year = "1990",
month = "1",
day = "1",
doi = "10.1128/MCB.10.8.3884",
language = "English (US)",
volume = "10",
pages = "3884--3895",
journal = "Molecular and Cellular Biology",
issn = "0270-7306",
publisher = "American Society for Microbiology",
number = "8",

}

TY - JOUR

T1 - A cis-acting element present in multiple genes serves as a repressor protein binding site for the yeast CAR1 gene

AU - Luche, R. M.

AU - Sumrada, R.

AU - Cooper, Terrance

PY - 1990/1/1

Y1 - 1990/1/1

N2 - Induction of the arginase (CAR1) gene expression in Saccharomyces cerevisiae has previously been shown to require participation of a cis-dominantly regulated upstream repression sequence (URS). Deletion of this element results in high-level expression of the CAR1 gene without inducer. To determine the structure of the CAR1 URS element, we performed a saturation mutagenesis. Results of the mutagenic analysis indicated that the CAR1 URS was a 9-base-pair palindromic sequence, 5'-AGCCGCCGA-3'. A DNA fragment containing this sequence was shown to bind one or more proteins by a gel shift assay. DNA fragments containing point mutations that completely eliminated URS function were not effective competitors in this assay, whereas those which supported URS function were effective competitors. Sequences in the 5'-flanking regions of 14 other genes were found to be homologous to the CAR1 URS. These sequences were shown to support varying degrees of URS function in the expression vector assay, to bind protein as demonstrated by the gel shift assay, and to compete with a DNA fragment containing the CAR 1 URS for protein binding. These results indicate that the CAR1 URS element possesses the characteristics of a repressor binding site. These results indicate that the CAR1 URS element possesses the characteristics of a repressor binding site. Further, they are consistent with the suggestion that sites homologous to the CAR1 URS may be situated in the 5'-flanking regions of multiple unrelated yeast genes. The widespread occurrence of this element raises the possibility that it is the target site for one or more negatively acting general transcription factors.

AB - Induction of the arginase (CAR1) gene expression in Saccharomyces cerevisiae has previously been shown to require participation of a cis-dominantly regulated upstream repression sequence (URS). Deletion of this element results in high-level expression of the CAR1 gene without inducer. To determine the structure of the CAR1 URS element, we performed a saturation mutagenesis. Results of the mutagenic analysis indicated that the CAR1 URS was a 9-base-pair palindromic sequence, 5'-AGCCGCCGA-3'. A DNA fragment containing this sequence was shown to bind one or more proteins by a gel shift assay. DNA fragments containing point mutations that completely eliminated URS function were not effective competitors in this assay, whereas those which supported URS function were effective competitors. Sequences in the 5'-flanking regions of 14 other genes were found to be homologous to the CAR1 URS. These sequences were shown to support varying degrees of URS function in the expression vector assay, to bind protein as demonstrated by the gel shift assay, and to compete with a DNA fragment containing the CAR 1 URS for protein binding. These results indicate that the CAR1 URS element possesses the characteristics of a repressor binding site. These results indicate that the CAR1 URS element possesses the characteristics of a repressor binding site. Further, they are consistent with the suggestion that sites homologous to the CAR1 URS may be situated in the 5'-flanking regions of multiple unrelated yeast genes. The widespread occurrence of this element raises the possibility that it is the target site for one or more negatively acting general transcription factors.

UR - http://www.scopus.com/inward/record.url?scp=0025345737&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025345737&partnerID=8YFLogxK

U2 - 10.1128/MCB.10.8.3884

DO - 10.1128/MCB.10.8.3884

M3 - Article

VL - 10

SP - 3884

EP - 3895

JO - Molecular and Cellular Biology

JF - Molecular and Cellular Biology

SN - 0270-7306

IS - 8

ER -