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PMID: 8633068 Published · ppublish English Comparative Study Journal Article

Differences in the RNA binding sites of iron regulatory proteins and potential target diversity.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 93 ·No. 9 ·1996-04-30 ·Pages 4345-9

Butt J, Kim HY, Basilion JP, Cohen S, Iwai K, Philpott CC, Altschul S, Klausner RD, Rouault TA

Abstract

Posttranscriptional regulation of genes of mammalian iron metabolism is mediated by the interaction of iron regulatory proteins (IRPs) with RNA stem-loop sequence elements known as iron-responsive elements (IREs). There are two identified IRPs, IRP1 and IRP2, each of which binds consensus IREs present in eukaryotic transcripts with equal affinity. Site-directed mutagenesis of IRP1 and IRP2 reveals that, although the binding affinities for consensus IREs are indistinguishable, the contributions of arginine residues in the active-site cleft to the binding affinity are different in the two RNA binding sites. Furthermore, although each IRP binds the consensus IRE with high affinity, each IRP also binds a unique alternative ligand, which was identified in an in vitro systematic evolution of ligands by exponential enrichment procedure. Differences in the two binding sites may be important in the function of the IRE-IRP regulatory system.

MeSH Terms
Alternative Splicing Amino Acid Sequence Animals Base Sequence Binding Sites Binding, Competitive Cell Line Chlorocebus aethiops Consensus Sequence DNA Primers Humans Iron Regulatory Protein 1 Iron Regulatory Protein 2 Iron-Regulatory Proteins Iron-Sulfur Proteins/biosynthesis,chemistry,metabolism Kinetics Molecular Sequence Data Nucleic Acid Conformation Point Mutation Polymerase Chain Reaction RNA, Messenger/chemistry,metabolism RNA-Binding Proteins/biosynthesis,chemistry,metabolism Recombinant Proteins/biosynthesis,chemistry,metabolism Transfection
Chemicals
DNA Primers Iron-Regulatory Proteins Iron-Sulfur Proteins RNA, Messenger RNA-Binding Proteins Recombinant Proteins Iron Regulatory Protein 1 Iron Regulatory Protein 2
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Butt J
Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA.
Kim H Y
Basilion J P
Cohen S
Iwai K
Philpott C C
Altschul S
Klausner R D
Rouault T A
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.93.9.4345
Published
1996-04-30
Pages
4345-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC39539
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.93.9.4345 PMC Full Text View journal details
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