Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1992 Dec;66(12):7438–7443. doi: 10.1128/jvi.66.12.7438-7443.1992

Distribution of conserved and specific epitopes on the VP8 subunit of rotavirus VP4.

G Larralde 1, M Gorziglia 1
PMCID: PMC240451  PMID: 1279204

Abstract

Three cDNA clones comprising the VP8 subunit of the VP4 of human rotavirus strain KU (VP7 serotype G1; VP4 serotype P1A) G1 were constructed. The corresponding encoded peptides were designated according to their locations in the VP8 subunit as A (amino acids 1 to 102), B (amino acids 84 to 180), and C (amino acids 150 to 246 plus amino acids 247 to 251 from VP5). In addition, cDNA clones encoding peptide B of the VP8 subunit of the VP4 gene from human rotavirus strains DS-1 (G2; P1B) and 1076 (G2; P2) were also constructed. These DNA fragments were inserted into plasmid pGEMEX-1 and expressed in Escherichia coli. Western immunoblot analysis using antisera to rotavirus strains KU (P1A), Wa (P1A), DS-1 (P1B), 1076 (P2), and M37 (P2) demonstrated that peptides A and C cross-reacted with heterotypic human rotavirus VP4 antisera, suggesting that these two peptides represent conserved epitopes in the VP8 subunit. In contrast, peptide B appears to be involved in the VP4 serotype and subtype specificities, because it reacted only with the corresponding serotype- and subtype-specific antiserum. Antiserum raised against peptide A, B, or C of strain KU contained a lower level of neutralizing activity than did that induced by the entire VP8 subunit. In addition, the serotype-specific neutralizing activity of anti-KU VP8 serum was ablated after adsorption with the KU VP8 protein but not with a mixture of peptides A, B, and C of strain KU, suggesting that most of the serotype-specific epitopes in the VP8 subunit are conformational and are dependent on the entire amino acid sequence of VP8.

Full text

PDF
7438

Images in this article

7439Image
on p.7439
7440Image
on p.7440
7440Image
on p.7440
7441Image
on p.7441
7441Image
on p.7441

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Browning G. F., Chalmers R. M., Fitzgerald T. A., Snodgrass D. R. Serological and genomic characterization of L338, a novel equine group A rotavirus G serotype. J Gen Virol. 1991 May;72(Pt 5):1059–1064. doi: 10.1099/0022-1317-72-5-1059. [DOI] [PubMed] [Google Scholar]
  2. Gorziglia M., Green K., Nishikawa K., Taniguchi K., Jones R., Kapikian A. Z., Chanock R. M. Sequence of the fourth gene of human rotaviruses recovered from asymptomatic or symptomatic infections. J Virol. 1988 Aug;62(8):2978–2984. doi: 10.1128/jvi.62.8.2978-2984.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Gorziglia M., Hoshino Y., Buckler-White A., Blumentals I., Glass R., Flores J., Kapikian A. Z., Chanock R. M. Conservation of amino acid sequence of VP8 and cleavage region of 84-kDa outer capsid protein among rotaviruses recovered from asymptomatic neonatal infection. Proc Natl Acad Sci U S A. 1986 Sep;83(18):7039–7043. doi: 10.1073/pnas.83.18.7039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gorziglia M., Larralde G., Kapikian A. Z., Chanock R. M. Antigenic relationships among human rotaviruses as determined by outer capsid protein VP4. Proc Natl Acad Sci U S A. 1990 Sep;87(18):7155–7159. doi: 10.1073/pnas.87.18.7155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hoshino Y., Sereno M. M., Midthun K., Flores J., Kapikian A. Z., Chanock R. M. Independent segregation of two antigenic specificities (VP3 and VP7) involved in neutralization of rotavirus infectivity. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8701–8704. doi: 10.1073/pnas.82.24.8701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hoshino Y., Wyatt R. G., Greenberg H. B., Flores J., Kapikian A. Z. Serotypic similarity and diversity of rotaviruses of mammalian and avian origin as studied by plaque-reduction neutralization. J Infect Dis. 1984 May;149(5):694–702. doi: 10.1093/infdis/149.5.694. [DOI] [PubMed] [Google Scholar]
  7. Larralde G., Flores J. Identification of gene 4 alleles among human rotaviruses by polymerase chain reaction-derived probes. Virology. 1990 Nov;179(1):469–473. doi: 10.1016/0042-6822(90)90317-k. [DOI] [PubMed] [Google Scholar]
  8. Larralde G., Li B. G., Kapikian A. Z., Gorziglia M. Serotype-specific epitope(s) present on the VP8 subunit of rotavirus VP4 protein. J Virol. 1991 Jun;65(6):3213–3218. doi: 10.1128/jvi.65.6.3213-3218.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Mackow E. R., Yamanaka M. Y., Dang M. N., Greenberg H. B. DNA amplification-restricted transcription-translation: rapid analysis of rhesus rotavirus neutralization sites. Proc Natl Acad Sci U S A. 1990 Jan;87(2):518–522. doi: 10.1073/pnas.87.2.518. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Offit P. A., Blavat G. Identification of the two rotavirus genes determining neutralization specificities. J Virol. 1986 Jan;57(1):376–378. doi: 10.1128/jvi.57.1.376-378.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Padilla-Noriega L., Fiore L., Rennels M. B., Losonsky G. A., Mackow E. R., Greenberg H. B. Humoral immune responses to VP4 and its cleavage products VP5* and VP8* in infants vaccinated with rhesus rotavirus. J Clin Microbiol. 1992 Jun;30(6):1392–1397. doi: 10.1128/jcm.30.6.1392-1397.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Prasad B. V., Wang G. J., Clerx J. P., Chiu W. Three-dimensional structure of rotavirus. J Mol Biol. 1988 Jan 20;199(2):269–275. doi: 10.1016/0022-2836(88)90313-0. [DOI] [PubMed] [Google Scholar]
  13. Qian Y., Green K. Y. Human rotavirus strain 69M has a unique VP4 as determined by amino acid sequence analysis. Virology. 1991 May;182(1):407–412. doi: 10.1016/0042-6822(91)90691-4. [DOI] [PubMed] [Google Scholar]
  14. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. doi: 10.1126/science.2999980. [DOI] [PubMed] [Google Scholar]
  15. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  16. Urasawa T., Urasawa S., Taniguchi K. Sequential passages of human rotavirus in MA-104 cells. Microbiol Immunol. 1981;25(10):1025–1035. doi: 10.1111/j.1348-0421.1981.tb00109.x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES