Nucleic Acids Research, 1993, Vol. 21, No. 23 5316-5322
© 1993
MOLECULAR BIOLOGY |
Translationaj repression by the human iron-regulatory factor (IRF) in Saccharomyces cerevisiae
Department of Gene Expression, Gesellschaft für Biotechnologische Forschung mbH (GBF) Mascheroder Weg 1, 38124 Braunschweig 1European Molecular Biology Laboratory (EMBL), Gene Expression Programme Meyerhofstrasse 1, 69117 Heidelberg, Germany
*To whom correspondence should be addressed
Received September 25, 1993. Revised October 25, 1993. Accepted October 25, 1993.
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.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  J. C. Canzoneri and A. K. Oyelere Interaction of anthracyclines with iron responsive element mRNAs Nucleic Acids Res., December 1, 2008; 36(21): 6825 - 6834. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. M. X. Hughes, M. Ptushkina, Md. M. Karim, N. Koloteva, T. von der Haar, and J. E. G. McCarthy Translational Repression by Human 4E-BP1 in Yeast Specifically Requires Human eIF4E as Target J. Biol. Chem., February 5, 1999; 274(6): 3261 - 3264. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Paraskeva, N. K. Gray, B. Schlager, K. Wehr, and M. W. Hentze Ribosomal Pausing and Scanning Arrest as Mechanisms of Translational Regulation from Cap-Distal Iron-Responsive Elements Mol. Cell. Biol., January 1, 1999; 19(1): 807 - 816. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. E. G. McCarthy Posttranscriptional Control of Gene Expression in Yeast Microbiol. Mol. Biol. Rev., December 1, 1998; 62(4): 1492 - 1553. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Paraskeva, A. Atzberger, and M. W. Hentze A translational repression assay procedure (TRAP) for RNA-protein interactions in vivo PNAS, February 3, 1998; 95(3): 951 - 956. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Koloteva, P. P. Muller, and J. E. G. McCarthy The Position Dependence of Translational Regulation via RNA-RNA and RNA-Protein Interactions in the 5'-Untranslated Region of Eukaryotic mRNA Is a Function of the Thermodynamic Competence of 40 S Ribosomes in Translational Initiation J. Biol. Chem., June 27, 1997; 272(26): 16531 - 16539. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Linz, N. Koloteva, S. Vasilescu, and J. E.G. McCarthy Disruption of Ribosomal Scanning on the 5'-Untranslated Region, and Not Restriction of Translational Initiation per se, Modulates the Stability of Nonaberrant mRNAs in the Yeast Saccharomyces cerevisiae J. Biol. Chem., April 4, 1997; 272(14): 9131 - 9140. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Wang and M. S. Sachs Arginine-specific Regulation Mediated by the Neurospora crassa arg-2 Upstream Open Reading Frame in a Homologous, Cell-free in Vitro Translation System J. Biol. Chem., January 3, 1997; 272(1): 255 - 261. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Vasilescu, M. Ptushkina, B. Linz, P. P. Müller, and J. E. G. McCarthy Mutants of Eukaryotic Initiation Factor eIF-4E with Altered mRNA Cap Binding Specificity Reprogram mRNA Selection by Ribosomes in Saccharomyces cerevisiae J. Biol. Chem., March 22, 1996; 271(12): 7030 - 7037. [Abstract] [Full Text] [PDF]
|
|