Arias, Carlos F. and López, Susana and Bell, John R. and Strauss, James H. (1984) Primary structure of the neutralization antigen of simian rotavirus SA11 as deduced from cDNA sequence. Journal of Virology, 50 (2). pp. 657-661. ISSN 0022-538X. PMCID PMC255701. http://resolver.caltech.edu/CaltechAUTHORS:ARIjvir84
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DNA sequences complementary to the double-stranded RNA coding for the neutralization antigen (glycoprotein VP7) of simian rotavirus SA11 have been cloned. The VP7 gene consists of 1,062 nucleotides, containing an uninterrupted coding sequence of 978 nucleotides which specifies a glycoprotein of 326 amino acids. The significance of a second possible initiation site 30 nucleotides downstream from the first is discussed. Partial amino acid sequence of this glycoprotein showed unequivocally that the cloned segment (segment 9) codes for glycoprotein VP7 of SA11. The resulting amino acid sequence contained only one carbohydrate acceptor site. Possible sites of membrane interaction and antigenic determinants are discussed based on the analysis of the hydrophobicity and hydrophilicity profiles of VP7.
|Additional Information:||Copyright © 1984, American Society for Microbiology. Received 18 July 1983/Accepted 28 October 1983 We are grateful to C. Rice for helpful discussions and advice on the sequencing and cloning experiments and to R. Espejo for critical discussion of the manuscript. We thank H. Huang for the gift of his recently developed plasmid pMT21. The computer programs used in this work were written by T. Hunkapiller, and we thank him for their use. This work was supported in part by Public Health Service grants GM 06965 and Al 10793 from the National Institutes of Health and grant PCM 8022830 from the National Science Foundation. C.A. and S.L. were supported by a grant from the Universidad Nacional Autónoma de Mexico. ADDENDUM: Just before this manuscript was submitted, the sequence of VP7 appeared (G. W. Both, J. S. Mattick, and A. R. Bellamy, Proc. Natl. Acad. Sci. U.S.A. 80:3091-3095, 1983). Our work confirms the coding assignments for VP7 by a different method, the main features of the nucleotide sequence of gene 9, and the inferred protein sequence. Our SAl was obtained from H. H. Malherbe in 1977, and significant divergence from other preparations might be expected after continuous passage for more than 5 years. However, comparison of the obtained sequences reveals that they have been highly conserved: only 12 nucleotides were found to differ in both strains. Seven of these mutations represented a change in an amino acid, and the other five were silent. Three of the amino acid changes were located in the hydrophobic regions proximal to the NH2 terminus, but all three were conservative. Only one of the other four amino acid changes is significative, involving nucleotide 389 and changing Gly-114 to Glu-114. None of the nucleotide mutations occurred at the two possible initiation sites. After submitting this manuscript, a closely related paper describing the nucleotide sequence of the gene encoding the neutralizing antigen of the bovine rotavirus UK appeared (T. C. Elleman, P. A. Hoyme, M. L. Dyall-Smith, I. H. Holmes, and A. A. Azad, Nucleic Acids Res. 11:4689-4701, 1983). The characteristics of the UK gene are strikingly similar to those reported here for the VP7 SA11 gene: it is 1,062 base pairs long, with a long open reading frame of 326 amino acids starting at the first of two in-phase possible initiation codons at nucleotide positions 49 and 136. The hydrophatic profile of the deduced protein showed two NH2-terminal hydrophobic regions and the presence of a carboxy-terminal highly hydrophilic region that may represent a major antigenic determinant. In addition to the one possible glycosylation site found in glycoprotein VP7 of simian rotavirus SA11 at amino acid 69, there are two more putative carbohydrate attachment sites in the bovine protein at amino acids 238 and 318. By comparing the UK and SA11 sequences, we found a 77% homology at the nucleotide level and a 86% homology at the amino acid level.|
|PubMed Central ID:||PMC255701|
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|Deposited On:||15 Mar 2006|
|Last Modified:||15 Dec 2015 22:56|
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