Abstract
The transport of messenger RNAs (mRNAs) from the nucleus to the cytoplasm involves adapter proteins that bind the mRNA as well as receptor proteins that interact with the nuclear pore complex. We demonstrate the utility of cell-permeable peptides designed to interfere with interactions between potential adapter and receptor proteins to define the pathways accessed by particular mRNAs. We show that HuR, a protein implicated in the stabilization of short-lived mRNAs containing AU-rich elements (AREs), serves as an adapter for c-fos mRNA export through two pathways. One involves the HuR shuttling domain, HNS, which exhibits a heat shock-sensitive interaction with transportin 2 (Trn2); the other involves two protein ligands of HuR-pp32 and APRIL-which contain leucine-rich nuclear export signals (NES) recognized by the export receptor CRM1. Heterokaryon and in situ hybridization experiments reveal that the peptides selectively block the nucleocytoplasmic shuttling of their respective adapter proteins without perturbing the overall cellular distribution of polyadenylated mRNAs.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Animals
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Antennapedia Homeodomain Protein
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Antigens, Surface*
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Biological Transport / drug effects
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Cell Line
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Cell Membrane Permeability*
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Cell Nucleus / drug effects
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Cell Nucleus / metabolism*
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Cytoplasm / drug effects
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Cytoplasm / metabolism*
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ELAV Proteins
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ELAV-Like Protein 1
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Exportin 1 Protein
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Genes, fos / genetics*
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Heat-Shock Response
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Homeodomain Proteins / chemistry
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Homeodomain Proteins / metabolism
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Humans
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Karyopherins / metabolism
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Molecular Sequence Data
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Neuropeptides / chemistry
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Neuropeptides / metabolism
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Nuclear Proteins / chemistry
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Nuclear Proteins / metabolism
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Peptide Fragments / metabolism*
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Peptide Fragments / pharmacology
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Phosphoproteins / metabolism
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Protein Binding / drug effects
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Protein Structure, Tertiary
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RNA Stability
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RNA, Messenger / genetics
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RNA, Messenger / metabolism*
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RNA-Binding Proteins / antagonists & inhibitors
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RNA-Binding Proteins / chemistry
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RNA-Binding Proteins / metabolism*
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Receptors, Cytoplasmic and Nuclear*
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Regulatory Sequences, Nucleic Acid / genetics
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Reproducibility of Results
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Tetrahydrofolate Dehydrogenase / genetics
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Transcription Factors*
Substances
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ANP32B protein, human
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Antennapedia Homeodomain Protein
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Antigens, Surface
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ELAV Proteins
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ELAV-Like Protein 1
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ELAVL1 protein, human
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Homeodomain Proteins
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Karyopherins
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Neuropeptides
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Nuclear Proteins
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Peptide Fragments
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Phosphoproteins
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RNA, Messenger
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RNA-Binding Proteins
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Receptors, Cytoplasmic and Nuclear
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Transcription Factors
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Tetrahydrofolate Dehydrogenase