A cytosolic protein binds to structural elements within the iron regulatory region of the transferrin receptor mRNA.
The level of mRNA encoding the transferrin receptor (TfR) is regulated by iron, and this regulation is mediated by a portion of the 3' untranslated region (UTR) of the TfR transcript. This portion of 3' UTR of the human TfR mRNA contains five RNA elements that have structural similarity to the iron-responsive element (IRE) found as a single copy in the 5' UTR of the mRNA for ferritin, whose translation is regulated by iron. Moreover, five very similar elements are also contained in the 3' UTR of the chicken TfR mRNA. Cytosolic extracts of human cell lines are shown by a gel shift assay involving RNase T1 protection to contain an IRE-binding protein capable of specific interaction with the human TfR 3' UTR. When the protecting protein is removed, the protected RNA can be digested with RNase T1 to yield oligoribonucleotide fragments characteristic of two of the IREs contained in the TfR 3' UTR. As judged by cross-competition experiments, the same IRE-binding protein interacts with the ferritin IRE. The apparent affinity of RNA sequence elements for the IRE-binding protein is shown to depend upon the sequence of the RNA. A comprehensive secondary structure for the regulatory region of the TfR mRNA is proposed based on the experimentally demonstrated presence of at least two IRE-like structural elements.
Koeller
D M
DM
Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, Bethesda, MD 20892.
Casey
J L
JL
Hentze
M W
MW
Gerhardt
E M
EM
Chan
L N
LN
Klausner
R D
RD
Harford
J B
JB
D001665:Binding Sites; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000502:physiology*; D003600:Cytosol / Q000502:physiology; D005293:Ferritins / Q000502:physiology; D006801:Humans; D006860:Hydrogen Bonding; D007501:Iron / Q000502:physiology*; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism* / Q000648:ultrastructure; D016601:RNA-Binding Proteins; D011990:Receptors, Transferrin / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid; D006163:Ribonuclease T1 / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D002352:Carrier Proteins; D009841:Oligonucleotides; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011990:Receptors, Transferrin; D005293:Ferritins; D007501:Iron; D006163:Ribonuclease T1