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PMID: 1854587 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Characteristics of the interaction of the ferritin repressor protein with the iron-responsive element.

Biology of metals ·Vol. 4 ·No. 1 ·1991-00-00 ·Pages 48-55

Swenson GR, Patino MM, Beck MM, Gaffield L, Walden WE

Abstract

The iron-responsive regulation of ferritin mRNA translation is mediated by the specific interaction of the ferritin repressor protein (FRP) with the iron-responsive element (IRE), a highly conserved 28-nucleotide sequence located in the 5' untranslated region of ferritin mRNAs. The IRE alone is necessary and sufficient to confer repression of translation by FRP upon a heterologous message, chloramphenicol acetyltransferase, in an in vitro translation system. The activity of FRP is sensitive to iron in vivo. Cytoplasmic extracts of rabbit kidney cells show reduction of FRP activity when grown in the presence of iron, as detected by RNA band shift assay. Using a nitrocellulose filter binding assay to examine the interaction of FRP with the IRE in more detail, we find that purified FRP has a single high-affinity binding site for the IRE with a Kd of 20-50 pM. Hemin pretreatment decreases the total amount of FRP which can bind to the IRE. This effect is dependent on hemin concentration. Interestingly, the FRP which remains active at a given hemin concentration binds to the IRE with the same high affinity as untreated FRP. A variety of hemin concentrations were examined for their effect on preformed FRP/IRE complexes. All hemin concentrations tested resulted in rapid complex breakdown. The final amount of complex breakdown corresponds to the concentration of hemin present in the reaction. The effect of hemin on FRP activity suggests that a specific hemin binding site exists on FRP.

MeSH Terms
Animals Carrier Proteins/drug effects,metabolism Diamide/pharmacology Ferritins/genetics,metabolism Hemin/pharmacology Humans Iron/metabolism Iron Regulatory Protein 1 Iron-Regulatory Proteins Kinetics Protein Biosynthesis RNA, Messenger/drug effects,genetics,metabolism Transcription, Genetic
Chemicals
Carrier Proteins Iron-Regulatory Proteins RNA, Messenger Diamide Hemin Ferritins Iron Iron Regulatory Protein 1
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Swenson G R
Department of Microbiology and Immunology, University of Illinois, Chicago 60612.
Patino M M
Beck M M
Gaffield L
Walden W E
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Article Info
Journal
Biology of metals
Abbr.
Biol Met
ISSN
0933-5854
DOI
10.1007/BF01135557
Published
1991-00-00
Pages
48-55
Language
English
Region
Germany
NLM ID
8915662
Subset
IM
External Links
PubMed source DOI: 10.1007/BF01135557
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