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PMID: 8265343 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Translational repression by the human iron-regulatory factor (IRF) in Saccharomyces cerevisiae.

Nucleic acids research ·Vol. 21 ·No. 23 ·1993-11-25 ·Pages 5316-22

Oliveira CC, Goossen B, Zanchin NI, McCarthy JE, Hentze MW, Stripecke R

Abstract

The regulation of the synthesis of ferritin and erythroid 5-aminolevulinate synthase in mammalian cells is mediated by the interaction of the iron regulatory factor (IRF) with a specific recognition site, the iron responsive element (IRE), in the 5' untranslated regions (UTRs) of the respective mRNAs. A new modular expression system was designed to allow reconstruction of this regulatory system in Saccharomyces cerevisiae. This comprised two components: a constitutively expressed reporter gene (luc; encoding luciferase) preceded by a 5' UTR including an IRE sequence, and an inducibly expressed cDNA encoding human IRF. Induction of the latter led to the in vivo synthesis of IRF, which in turn showed IRE-binding activity and also repressed translation of the luc mRNA bearing an IRE-containing 5' UTR. The upper stem-loop region of an IRE, with no further IRE-specific flanking sequences, sufficed for recognition and repression by IRF. Translational regulation of IRE-bearing mRNAs could also be demonstrated in cell-free yeast extracts. This work defines a minimal system for IRF/IRE translational regulation in yeast that requires no additional mammalian-specific components, thus providing direct proof that IRF functions as a translational repressor in vivo. It should be a useful tool as the basis for more detailed studies of eukaryotic translational regulation.

MeSH Terms
Base Sequence Gene Expression Regulation Gene Expression Regulation, Fungal Humans Iron-Regulatory Proteins Molecular Sequence Data Protein Biosynthesis RNA, Messenger/genetics RNA-Binding Proteins/genetics Recombinant Proteins Repressor Proteins/genetics Saccharomyces cerevisiae/genetics Transcription, Genetic
Chemicals
Iron-Regulatory Proteins RNA, Messenger RNA-Binding Proteins Recombinant Proteins Repressor Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Oliveira C C
Department of Gene Expression, Gesellschaft für Biotechnologische Forschung mbH (GBF), Braunschweig, Germany.
Goossen B
Zanchin N I
McCarthy J E
Hentze M W
Stripecke R
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/21.23.5316
Published
1993-11-25
Pages
5316-22
Language
English
Region
England
NLM ID
0411011
PMCID
PMC310564
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/21.23.5316 PMC Full Text View journal details
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