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PMID: 1314953 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Regulation of yeast COX6 by the general transcription factor ABF1 and separate HAP2- and heme-responsive elements.

Molecular and cellular biology ·Vol. 12 ·No. 5 ·1992-05-00 ·Pages 2302-14

Trawick JD, Kraut N, Simon FR, Poyton RO

Abstract

Transcription of the Saccharomyces cerevisiae COX6 gene is regulated by heme and carbon source. It is also affected by the HAP2/3/4 transcription factor complex and by SNF1 and SSN6. Previously, we have shown that most of this regulation is mediated through UAS6, an 84-bp upstream activation segment of the COX6 promoter. In this study, by using linker scanning mutagenesis and protein binding assays, we have identified three elements within UAS6 and one element downstream of it that are important. Two of these, HDS1 (heme-dependent site 1; between -269 and -251 bp) and HDS2 (between -228 and -220 bp), mediate regulation of COX6 by heme. Both act negatively. The other two elements, domain 2 (between -279 and -269 bp) and domain 1 (between -302 and -281 bp), act positively. Domain 2 is required for optimal transcription in cells grown in repressing but not derepressing carbon sources. Domain 1 is essential for transcription per se in cells grown on repressing carbon sources, is required for optimal transcription in cells grown on a derepressing carbon source, is sufficient for glucose repression-derepression, and is the element of UAS6 at which HAP2 affects COX6 transcription. This element contains the major protein binding sites within UAS6. It has consensus binding sequences for ABF1 and HAP2. Gel mobility shift experiments show that domain 1 binds ABF1 and forms different numbers of DNA-protein complexes in extracts from cells grown in repressing or derepressing carbon sources. In contrast, gel mobility shift experiments have failed to reveal that HAP2 or HAP3 binds to domain 1 or that hap3 mutations affect the complexes bound to it. Together, these findings permit the following conclusions: COX6 transcription is regulated both positively and negatively; heme and carbon source exert their effects through different sites; domain 1 is absolutely essential for transcription on repressing carbon sources; ABF1 is a major component in the regulation of COX6 transcription; and the HAP2/3/4 complex most likely affects COX6 transcription indirectly.

Related Genes
COX6 HAP2 HAP3 HAP4 lacZ
MeSH Terms
Base Sequence CCAAT-Binding Factor DNA, Fungal/genetics,metabolism DNA-Binding Proteins Electron Transport Complex IV/genetics Fungal Proteins/genetics,metabolism Gene Expression Regulation, Enzymologic/drug effects Gene Expression Regulation, Fungal/drug effects Heme/pharmacology Kinetics Molecular Sequence Data Mutagenesis, Insertional Oligodeoxyribonucleotides Plasmids Promoter Regions, Genetic Recombinant Fusion Proteins/metabolism Saccharomyces cerevisiae/enzymology,genetics Saccharomyces cerevisiae Proteins Transcription Factors/genetics,metabolism Transcription, Genetic/drug effects beta-Galactosidase/genetics,metabolism
Chemicals
ABF1 protein, S cerevisiae CCAAT-Binding Factor DNA, Fungal DNA-Binding Proteins Fungal Proteins Oligodeoxyribonucleotides Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins Transcription Factors Heme Electron Transport Complex IV beta-Galactosidase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Trawick J D
Hepatobiliary Research Center, University of Colorado Health Sciences Center, Denver 80262.
Kraut N
Simon F R
Poyton R O
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54 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.12.5.2302-2314.1992
Published
1992-05-00
Pages
2302-14
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC364402
Subset
IM
Grants
NIGMS NIH HHS · GM 34827 · United States
NIGMS NIH HHS · GM 39324 · United States
NIGMS NIH HHS · GM30228 · United States
External Links
PubMed source DOI: 10.1128/mcb.12.5.2302-2314.1992 PMC Full Text View journal details
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