Two proline residues are essential in the calcium-binding activity of rotavirus VP7 outer capsid protein.
Rotavirus maturation and stability of the outer capsid are calcium-dependent processes. It has been shown previously that the concentration of Ca2+-solubilizing outer capsid proteins from rotavirus particles is dependent on the virus strain. This property of viral particles has been associated with the gene coding for VP7 (gene 9). In this study the correlation between VP7 and resistance to low [Ca2+] was confirmed by analyzing the origin of gene 9 from reassortant viruses prepared under the selective pressure of low [Ca2+]. After chemical mutagenesis, we selected mutant viruses of the bovine strain RF that are more resistant to low [Ca2+]. The genes coding for the VP7 proteins of these independent mutants have been sequenced. Sequence analysis confirmed that these mutants are independent and revealed that all mutant VP7 proteins have proline 75 changed to leucine and have an outer capsid that solubilized at low [Ca2+]. The mutation of proline 279 to serine is found in all but two mutants. The phenotype of mutants having a single proline change can be distinguished from the phenotype of mutants having two proline changes. Sequence analysis showed that position 75 is in a region (amino acids 65 to 78) of great variability and that proline 75 is present in most of the bovine strains. In contrast, proline 279 is in a conserved region and is conserved in all the VP7 sequences in data banks. This region is rich in oxygenated residues that are correctly allocated in the metal-coordinating positions of the Ca2+-binding EF-hand structure pattern, suggesting that this region is important in the Ca2+ binding of VP7.
Gajardo
R
R
Laboratoire de Virologie et Immunologie Moléculaires, INRA, C.R.J., Domaine de Vilvert, France.
Vende
P
P
Poncet
D
D
Cohen
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000956:Antigens, Viral; D002118:Calcium / Q000378:metabolism* / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002213:Capsid / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D036022:Capsid Proteins; D002417:Cattle; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011392:Proline; D012401:Rotavirus / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism
D000956:Antigens, Viral; D002135:Calcium-Binding Proteins; D036022:Capsid Proteins; C050779:VP7 protein, Rotavirus; D011392:Proline; D002118:Calcium