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. 1996 Nov;70(11):7833–7841. doi: 10.1128/jvi.70.11.7833-7841.1996

The 3'-terminal consensus sequence of rotavirus mRNA is the minimal promoter of negative-strand RNA synthesis.

M J Wentz 1, J T Patton 1, R F Ramig 1
PMCID: PMC190854  PMID: 8892905

Abstract

We used an in vitro template-dependent replicase assay (D. Chen, C. Zeng, M. Wentz, M. Gorziglia, M. Estes, and R. Ramig. J. Virol. 68:7030-7039, 1994) to identify the cis-acting signals required for replication of a genome segment 9 template from the group A rotavirus strain OSU. The replicase phenotypes for a panel of templates with internal deletions or 3'-terminal truncations indicated that no essential replication signals were present within the open reading frame and that key elements were present in the 5' and 3' noncoding regions. Chimeric constructs containing portions of viral sequence ligated to a nonviral backbone were generated to further map the regions required for in vitro replication of segment 9. The data from these constructs showed that the 3'-terminal seven nucleotides of the segment 9 mRNA provided the minimum requirement for replication (minimal promoter). Analysis of additional chimeric templates demonstrated that sequences capable of enhancing replication from the minimal promoter were located immediately upstream of the minimal promoter and at the extreme 5' terminus of the template. Mutational analysis of the minimal promoter revealed that the 3'-terminal -CC residues are required for efficient replication. Comparison of the replication levels for templates with guanosines and uridines at nucleotides -4 to -6 from the 3' terminus compared with levels for templates containing neither of these residues at these positions indicated that either or both residues must be present in this region for efficient replication in vitro.

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Selected References

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  1. Ball L. A. Cellular expression of a functional nodavirus RNA replicon from vaccinia virus vectors. J Virol. 1992 Apr;66(4):2335–2345. doi: 10.1128/jvi.66.4.2335-2345.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bican P., Cohen J., Charpilienne A., Scherrer R. Purification and characterization of bovine rotavirus cores. J Virol. 1982 Sep;43(3):1113–1117. doi: 10.1128/jvi.43.3.1113-1117.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chen D. Y., Ramig R. F. Determinants of rotavirus stability and density during CsCl purification. Virology. 1992 Jan;186(1):228–237. doi: 10.1016/0042-6822(92)90077-3. [DOI] [PubMed] [Google Scholar]
  4. Chen D., Ramig R. F. Rescue of infectivity by in vitro transcapsidation of rotavirus single-shelled particles. Virology. 1993 Feb;192(2):422–429. doi: 10.1006/viro.1993.1057. [DOI] [PubMed] [Google Scholar]
  5. Chen D., Zeng C. Q., Wentz M. J., Gorziglia M., Estes M. K., Ramig R. F. Template-dependent, in vitro replication of rotavirus RNA. J Virol. 1994 Nov;68(11):7030–7039. doi: 10.1128/jvi.68.11.7030-7039.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cohen J., Laporte J., Charpilienne A., Scherrer R. Activation of rotavirus RNA polymerase by calcium chelation. Arch Virol. 1979;60(3-4):177–186. doi: 10.1007/BF01317489. [DOI] [PubMed] [Google Scholar]
  7. Desselberger U., Racaniello V. R., Zazra J. J., Palese P. The 3' and 5'-terminal sequences of influenza A, B and C virus RNA segments are highly conserved and show partial inverted complementarity. Gene. 1980 Feb;8(3):315–328. doi: 10.1016/0378-1119(80)90007-4. [DOI] [PubMed] [Google Scholar]
  8. Fodor E., Pritlove D. C., Brownlee G. G. The influenza virus panhandle is involved in the initiation of transcription. J Virol. 1994 Jun;68(6):4092–4096. doi: 10.1128/jvi.68.6.4092-4096.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Frilander M., Gottlieb P., Strassman J., Bamford D. H., Mindich L. Dependence of minus-strand synthesis on complete genomic packaging in the double-stranded RNA bacteriophage phi 6. J Virol. 1992 Aug;66(8):5013–5017. doi: 10.1128/jvi.66.8.5013-5017.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fujimura T., Esteban R., Esteban L. M., Wickner R. B. Portable encapsidation signal of the L-A double-stranded RNA virus of S. cerevisiae. Cell. 1990 Aug 24;62(4):819–828. doi: 10.1016/0092-8674(90)90125-x. [DOI] [PubMed] [Google Scholar]
  11. Gallegos C. O., Patton J. T. Characterization of rotavirus replication intermediates: a model for the assembly of single-shelled particles. Virology. 1989 Oct;172(2):616–627. doi: 10.1016/0042-6822(89)90204-3. [DOI] [PubMed] [Google Scholar]
  12. Gottlieb P., Qiao X., Strassman J., Frilander M., Mindich L. Identification of the packaging regions within the genomic RNA segments of bacteriophage phi 6. Virology. 1994 Apr;200(1):42–47. doi: 10.1006/viro.1994.1160. [DOI] [PubMed] [Google Scholar]
  13. Gottlieb P., Strassman J., Qiao X., Frilander M., Frucht A., Mindich L. In vitro packaging and replication of individual genomic segments of bacteriophage phi 6 RNA. J Virol. 1992 May;66(5):2611–2616. doi: 10.1128/jvi.66.5.2611-2616.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hagen M., Chung T. D., Butcher J. A., Krystal M. Recombinant influenza virus polymerase: requirement of both 5' and 3' viral ends for endonuclease activity. J Virol. 1994 Mar;68(3):1509–1515. doi: 10.1128/jvi.68.3.1509-1515.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Imai M., Akatani K., Ikegami N., Furuichi Y. Capped and conserved terminal structures in human rotavirus genome double-stranded RNA segments. J Virol. 1983 Jul;47(1):125–136. doi: 10.1128/jvi.47.1.125-136.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Li X., Palese P. Mutational analysis of the promoter required for influenza virus virion RNA synthesis. J Virol. 1992 Jul;66(7):4331–4338. doi: 10.1128/jvi.66.7.4331-4338.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Luo G. X., Luytjes W., Enami M., Palese P. The polyadenylation signal of influenza virus RNA involves a stretch of uridines followed by the RNA duplex of the panhandle structure. J Virol. 1991 Jun;65(6):2861–2867. doi: 10.1128/jvi.65.6.2861-2867.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mattion N. M., Cohen J., Aponte C., Estes M. K. Characterization of an oligomerization domain and RNA-binding properties on rotavirus nonstructural protein NS34. Virology. 1992 Sep;190(1):68–83. doi: 10.1016/0042-6822(92)91193-x. [DOI] [PubMed] [Google Scholar]
  20. McCrae M. A., McCorquodale J. G. Molecular biology of rotaviruses. V. Terminal structure of viral RNA species. Virology. 1983 Apr 15;126(1):204–212. doi: 10.1016/0042-6822(83)90472-5. [DOI] [PubMed] [Google Scholar]
  21. Pattnaik A. K., Ball L. A., LeGrone A., Wertz G. W. The termini of VSV DI particle RNAs are sufficient to signal RNA encapsidation, replication, and budding to generate infectious particles. Virology. 1995 Jan 10;206(1):760–764. doi: 10.1016/S0042-6822(95)80005-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Patton J. T. Synthesis of simian rotavirus SA11 double-stranded RNA in a cell-free system. Virus Res. 1986 Dec;6(3):217–233. doi: 10.1016/0168-1702(86)90071-7. [DOI] [PubMed] [Google Scholar]
  23. Patton J. T., Wentz M., Xiaobo J., Ramig R. F. cis-Acting signals that promote genome replication in rotavirus mRNA. J Virol. 1996 Jun;70(6):3961–3971. doi: 10.1128/jvi.70.6.3961-3971.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Petrie B. L., Graham D. Y., Hanssen H., Estes M. K. Localization of rotavirus antigens in infected cells by ultrastructural immunocytochemistry. J Gen Virol. 1982 Dec;63(2):457–467. doi: 10.1099/0022-1317-63-2-457. [DOI] [PubMed] [Google Scholar]
  25. Poncet D., Aponte C., Cohen J. Rotavirus protein NSP3 (NS34) is bound to the 3' end consensus sequence of viral mRNAs in infected cells. J Virol. 1993 Jun;67(6):3159–3165. doi: 10.1128/jvi.67.6.3159-3165.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Poncet D., Laurent S., Cohen J. Four nucleotides are the minimal requirement for RNA recognition by rotavirus non-structural protein NSP3. EMBO J. 1994 Sep 1;13(17):4165–4173. doi: 10.1002/j.1460-2075.1994.tb06734.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Sandino A. M., Jashes M., Faúndez G., Spencer E. Role of the inner protein capsid on in vitro human rotavirus transcription. J Virol. 1986 Nov;60(2):797–802. doi: 10.1128/jvi.60.2.797-802.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Shaw A. L., Rothnagel R., Chen D., Ramig R. F., Chiu W., Prasad B. V. Three-dimensional visualization of the rotavirus hemagglutinin structure. Cell. 1993 Aug 27;74(4):693–701. doi: 10.1016/0092-8674(93)90516-S. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wickner R. B. Double-stranded RNA virus replication and packaging. J Biol Chem. 1993 Feb 25;268(6):3797–3800. [PubMed] [Google Scholar]

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