Abstract
Rotavirus NSP4 plays multiple roles in viral pathogenesis, morphogenesis and replication. We previously reported a direct interaction between full-length NSP4 and the enterotoxic peptide composed of NSP4 residues 114-135 with full-length caveolin-1, the structural protein of caveolae. Caveolin-1 forms a hairpin loop in the cytoplasmic leaflet of plasma membrane caveolae. This unique orientation results in both termini of caveolin-1 exposed to the cytoplasm. The goal of this study was to map the caveolin-1 residues that interact with NSP4 to obtain a more complete picture of this binding event. Utilizing reverse yeast two-hybrid analyses and direct peptide binding assays, the NSP4 binding site was localized to caveolin-1 residues 2-22 and 161-178, at the amino- and carboxyl-termini, respectively. However, NSP4 binding to one of the termini was sufficient for the interaction.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Amino Acid Sequence
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Base Sequence
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Binding Sites
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Caveolin 1 / chemistry*
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Caveolin 1 / genetics
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Caveolin 1 / metabolism*
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DNA Primers / genetics
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Glycoproteins / chemistry
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Glycoproteins / genetics
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Glycoproteins / physiology*
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Humans
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In Vitro Techniques
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Molecular Sequence Data
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Peptide Fragments / chemistry
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Peptide Fragments / genetics
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Peptide Fragments / metabolism
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Protein Binding
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Protein Structure, Tertiary
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Rotavirus / genetics
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Rotavirus / pathogenicity*
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Rotavirus / physiology*
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Sequence Deletion
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Toxins, Biological / chemistry
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Toxins, Biological / genetics
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Toxins, Biological / physiology*
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Two-Hybrid System Techniques
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Viral Nonstructural Proteins / chemistry
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Viral Nonstructural Proteins / genetics
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Viral Nonstructural Proteins / physiology*
Substances
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CAV1 protein, human
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Caveolin 1
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DNA Primers
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Glycoproteins
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NS28 protein, rotavirus
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Peptide Fragments
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Recombinant Proteins
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Toxins, Biological
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Viral Nonstructural Proteins