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PMID: 15914670 Published · epublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

The ELAV RNA-stability factor HuR binds the 5'-untranslated region of the human IGF-IR transcript and differentially represses cap-dependent and IRES-mediated translation.

Nucleic acids research ·Vol. 33 ·No. 9 ·2005-00-00 ·Pages 2962-79

Meng Z, King PH, Nabors LB, Jackson NL, Chen CY, Emanuel PD, Blume SW

Abstract

The type I insulin-like growth factor receptor (IGF-IR) is an integral component in the control of cell proliferation, differentiation and apoptosis. The IGF-IR mRNA contains an extraordinarily long (1038 nt) 5'-untranslated region (5'-UTR), and we have characterized a diverse series of proteins interacting with this RNA sequence which may provide for intricate regulation of IGF-IR gene expression at the translational level. Here, we report the purification and identification of one of these IGF-IR 5'-UTR-binding proteins as HuR, using a novel RNA crosslinking/RNase elution strategy. Because HuR has been predominantly characterized as a 3'-UTR-binding protein, enhancing mRNA stability and generally increasing gene expression, we sought to determine whether HuR might serve a different function in the context of its binding the IGF-IR 5'-UTR. We found that HuR consistently repressed translation initiation through the IGF-IR 5'-UTR. The inhibition of translation by HuR was concentration dependent, and could be reversed in trans by addition of a fragment of the IGF-IR 5'-UTR containing the HuR binding sites as a specific competitor, or abrogated by deletion of the third RNA recognition motif of HuR. We determined that HuR repressed translation initiation through the IGF-IR 5'-UTR in cells as well, and that siRNA knockdown of HuR markedly increased IGF-IR protein levels. Interestingly, we also found that HuR potently inhibited IGF-IR translation mediated through internal ribosome entry. Kinetic assays were performed to investigate the mechanism of translation repression by HuR and the dynamic interplay between HuR and the translation apparatus. We found that HuR, occupying a cap-distal position, significantly delayed translation initiation mediated by cap-dependent scanning, but was eventually displaced from its binding site, directly or indirectly, as a consequence of ribosomal scanning. However, HuR perpetually blocked the activity of the IGF-IR IRES, apparently arresting the IRES-associated translation pre-initiation complex in an inactive state. This function of HuR as a 5'-UTR-binding protein and dual-purpose translation repressor may be critical for the precise regulation of IGF-IR expression essential to normal cellular homeostasis.

MeSH Terms
5' Untranslated Regions/metabolism Antigens, Surface/isolation & purification,metabolism Binding Sites Cell Line ELAV Proteins ELAV-Like Protein 1 Gene Expression Regulation Humans Protein Biosynthesis RNA Caps/metabolism RNA-Binding Proteins/antagonists & inhibitors,isolation & purification,metabolism Receptor, IGF Type 1/chemistry,genetics,metabolism Ribosomes/metabolism
Chemicals
5' Untranslated Regions Antigens, Surface ELAV Proteins ELAV-Like Protein 1 ELAVL1 protein, human RNA Caps RNA-Binding Proteins Receptor, IGF Type 1
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Meng Zheng
Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham Birmingham, AL 35294, USA.
King Peter H
Nabors L Burt
Jackson Nateka L
Chen Ching-Yi
Emanuel Peter D
Blume Scott W
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gki603
Published
2005-00-00
Epub
2005-00-24
Pages
2962-79
Language
English
Region
England
NLM ID
0411011
PMCID
PMC1140080
Subset
IM
Grants
NCI NIH HHS · K24 CA080916 · United States
NCI NIH HHS · R01 CA108886 · United States
NCI NIH HHS · CA108886 · United States
NCI NIH HHS · CA80916 · United States
External Links
PubMed source DOI: 10.1093/nar/gki603 PMC Full Text View journal details
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