Molecular characterization of the rotavirus NSP4 enterotoxin homologue from group B rotavirus.
The RNA segment (Gene 10) from a human group B rotavirus which encodes the homologue of the rotavirus enterotoxin (NSP4) has been cloned and sequenced. The gene is of the same length (751 nucleotides) as its better-characterized group A rotavirus counterpart but shows minimal homology (approximately 10%) to it at the primary sequence level. Despite this low level of sequence homology, secondary structure predictions for the group B protein (ADRV-NSP4) showed a close similarity of structural features with the group A protein. Full-length ADRV-NSP4 was expressed in Escherichia coli with an amino terminal 6xHis tag that was used to purify it to homogeneity. The cytotoxicity of the purified protein was examined in a rapid dye-uptake assay that assesses membrane permeability and was found to be comparable to its group A counterpart.
Guzman
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Department of Biological Sciences, University of Warwick, Gibbet Hill Rd, Coventry CV4 7AL, UK.
McCrae
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MA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000633:toxicity*; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000633:toxicity; D012401:Rotavirus / Q000235:genetics*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014118:Toxins, Biological / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000633:toxicity*; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000633:toxicity*
D006023:Glycoproteins; C081369:NS28 protein, rotavirus; D011994:Recombinant Proteins; D014118:Toxins, Biological; D017361:Viral Nonstructural Proteins