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PMID: 8602150 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The multifunctional transcription factors Abf1p, Rap1p and Reb1p are required for full transcriptional activation of the chromosomal PGK gene in Saccharomyces cerevisiae.

Molecular & general genetics : MGG ·Vol. 250 ·No. 3 ·1996-02-25 ·Pages 348-56

Packham EA, Graham IR, Chambers A

Abstract

We have identified two new transcription factor binding sites upstream of the previously defined UAS within the phosphoglycerate kinase (PGK) gene promoter in Saccharomyces cerevisiae. These sites are bound in vitro by the multifunctional factors Cpf1p and Reb1p. We have generated targeted deletions of Rap1p, Abf1p and Reb1p binding sites in the promoter of the chromosomal copy of the PGK gene. Northern blot analysis confirmed that most PGK promoter activity is mediated through the Rap1p binding site. However, significant effects are also mediated through both the Reb1p and Abf1p sites. In contrast, when the promoter is present on a high-copy-number plasmid, both the Abf1p and Reb1p sites play no role in transcriptional activation. The role of Cpf1p was examined using a cpf1 null strain. Cpf1p was found to have little if any, effect on activation of either the chromosomal or plasmid-borne PGK gene.

MeSH Terms
Base Sequence Basic Helix-Loop-Helix Leucine Zipper Transcription Factors Binding Sites Blotting, Northern Cloning, Molecular DNA Primers/chemistry DNA-Binding Proteins/genetics,metabolism,physiology Electrophoresis, Polyacrylamide Gel Fungal Proteins/genetics,metabolism,physiology GTP-Binding Proteins/genetics,metabolism Gene Expression Regulation, Fungal/genetics Molecular Sequence Data Phosphoglycerate Kinase/genetics,metabolism Promoter Regions, Genetic/genetics Saccharomyces cerevisiae/enzymology,genetics Saccharomyces cerevisiae Proteins Sequence Deletion/genetics Transcription Factors/genetics,metabolism Transcription, Genetic/genetics Transcriptional Activation rap GTP-Binding Proteins
Chemicals
ABF1 protein, S cerevisiae Basic Helix-Loop-Helix Leucine Zipper Transcription Factors CBF1 protein, S cerevisiae DNA Primers DNA-Binding Proteins Fungal Proteins REB1 protein, S cerevisiae Saccharomyces cerevisiae Proteins Transcription Factors Phosphoglycerate Kinase GTP-Binding Proteins rap GTP-Binding Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Packham E A
Department of Genetics, University of Nottingham, Queen's Medical Centre, Nottingham, UK.
Graham I R
Chambers A
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF02174393
Published
1996-02-25
Pages
348-56
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
External Links
PubMed source DOI: 10.1007/BF02174393
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