Rotavirus NSP4 interacts with both the amino- and carboxyl-termini of caveolin-1.
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.
Mir
Kiran D
KD
Department of Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX 77843, USA.
Parr
Rebecca D
RD
Schroeder
Friedhelm
F
Ball
Judith M
JM
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D051242:Caveolin 1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D006801:Humans; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012401:Rotavirus / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology*; D017384:Sequence Deletion; D014118:Toxins, Biological / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D020798:Two-Hybrid System Techniques; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*
C495540:CAV1 protein, human; D051242:Caveolin 1; D017931:DNA Primers; D006023:Glycoproteins; C081369:NS28 protein, rotavirus; D010446:Peptide Fragments; D011994:Recombinant Proteins; D014118:Toxins, Biological; D017361:Viral Nonstructural Proteins