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. 1996 Mar;70(3):1941–1952. doi: 10.1128/jvi.70.3.1941-1952.1996

Capsid coding sequence is required for efficient replication of human rhinovirus 14 RNA.

K L McKnight 1, S M Lemon 1
PMCID: PMC190023  PMID: 8627720

Abstract

Mechanisms by which the plus-sense RNA genomes of picornaviruses are replicated remain poorly defined, but existing models do not suggest a role for sequences encoding the capsid proteins. However, candidate RNA replicons (delta P1 beta gal and delta P1Luc), representing the sequence of human rhinovirus 14 virus (HRV-14) with reporter protein sequences (beta-galactosidase or luciferase, respectively) replacing most of the P1 capsid-coding region, failed to replicate in transfected H1-HeLa cells despite efficient primary cleavage of the polyprotein. To determine which P1 sequences might be required for RNA replication, HRV-14 mutants in which segments of the P1 region were removed to frame from the genome were constructed. Mutants with deletions involving the 5'proximal 1,489 nucleotides of the P1 region replicated efficiently, while those with deletions involving the 3' 1,079 nucleotides did not. Reintroduction of the 3' P1 sequence into the nonreplicating delta P1Luc construct resulted in a new candidate replicon, delta P1Luc/VP3, which replicated well and expressed luciferase efficiently. Capsid proteins provided in trans by helper virus failed to rescue the nonreplicating delta P1Luc genome but were able to package the larger-than-genome-length delta P1Luc/VP3 replicon. Thus, a 3'-distal P1 capsid-coding sequence has a previously unrecognized cis-active function related to replication of HRV-14 RNA.

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Selected References

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  1. Alsaadi S., Hassard S., Stanway G. Sequences in the 5' non-coding region of human rhinovirus 14 RNA that affect in vitro translation. J Gen Virol. 1989 Oct;70(Pt 10):2799–2804. doi: 10.1099/0022-1317-70-10-2799. [DOI] [PubMed] [Google Scholar]
  2. Alvey J. C., Wyckoff E. E., Yu S. F., Lloyd R., Ehrenfeld E. cis- and trans-cleavage activities of poliovirus 2A protease expressed in Escherichia coli. J Virol. 1991 Nov;65(11):6077–6083. doi: 10.1128/jvi.65.11.6077-6083.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Andino R., Rieckhof G. E., Achacoso P. L., Baltimore D. Poliovirus RNA synthesis utilizes an RNP complex formed around the 5'-end of viral RNA. EMBO J. 1993 Sep;12(9):3587–3598. doi: 10.1002/j.1460-2075.1993.tb06032.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Andino R., Silvera D., Suggett S. D., Achacoso P. L., Miller C. J., Baltimore D., Feinberg M. B. Engineering poliovirus as a vaccine vector for the expression of diverse antigens. Science. 1994 Sep 2;265(5177):1448–1451. doi: 10.1126/science.8073288. [DOI] [PubMed] [Google Scholar]
  5. Barrera I., Schuppli D., Sogo J. M., Weber H. Different mechanisms of recognition of bacteriophage Q beta plus and minus strand RNAs by Q beta replicase. J Mol Biol. 1993 Jul 20;232(2):512–521. doi: 10.1006/jmbi.1993.1407. [DOI] [PubMed] [Google Scholar]
  6. Blair W. S., Semler B. L. Role for the P4 amino acid residue in substrate utilization by the poliovirus 3CD proteinase. J Virol. 1991 Nov;65(11):6111–6123. doi: 10.1128/jvi.65.11.6111-6123.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brown E. A., Day S. P., Jansen R. W., Lemon S. M. The 5' nontranslated region of hepatitis A virus RNA: secondary structure and elements required for translation in vitro. J Virol. 1991 Nov;65(11):5828–5838. doi: 10.1128/jvi.65.11.5828-5838.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Callahan P. L., Mizutani S., Colonno R. J. Molecular cloning and complete sequence determination of RNA genome of human rhinovirus type 14. Proc Natl Acad Sci U S A. 1985 Feb;82(3):732–736. doi: 10.1073/pnas.82.3.732. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Choi W. S., Pal-Ghosh R., Morrow C. D. Expression of human immunodeficiency virus type 1 (HIV-1) gag, pol, and env proteins from chimeric HIV-1-poliovirus minireplicons. J Virol. 1991 Jun;65(6):2875–2883. doi: 10.1128/jvi.65.6.2875-2883.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cole C. N., Baltimore D. Defective interfering particles of poliovirus. II. Nature of the defect. J Mol Biol. 1973 May 25;76(3):325–343. doi: 10.1016/0022-2836(73)90508-1. [DOI] [PubMed] [Google Scholar]
  11. Cole C. N. Defective interfering (di) particles of poliovirus. Prog Med Virol. 1975;20:180–207. [PubMed] [Google Scholar]
  12. Cole C. N., Smoler D., Wimmer E., Baltimore D. Defective interfering particles of poliovirus. I. Isolation and physical properties. J Virol. 1971 Apr;7(4):478–485. doi: 10.1128/jvi.7.4.478-485.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Collis P. S., O'Donnell B. J., Barton D. J., Rogers J. A., Flanegan J. B. Replication of poliovirus RNA and subgenomic RNA transcripts in transfected cells. J Virol. 1992 Nov;66(11):6480–6488. doi: 10.1128/jvi.66.11.6480-6488.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dewalt P. G., Lawson M. A., Colonno R. J., Semler B. L. Chimeric picornavirus polyproteins demonstrate a common 3C proteinase substrate specificity. J Virol. 1989 Aug;63(8):3444–3452. doi: 10.1128/jvi.63.8.3444-3452.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Duechler M., Skern T., Blaas D., Berger B., Sommergruber W., Kuechler E. Human rhinovirus serotype 2: in vitro synthesis of an infectious RNA. Virology. 1989 Jan;168(1):159–161. doi: 10.1016/0042-6822(89)90414-5. [DOI] [PubMed] [Google Scholar]
  16. French R., Ahlquist P. Intercistronic as well as terminal sequences are required for efficient amplification of brome mosaic virus RNA3. J Virol. 1987 May;61(5):1457–1465. doi: 10.1128/jvi.61.5.1457-1465.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hagino-Yamagishi K., Nomoto A. In vitro construction of poliovirus defective interfering particles. J Virol. 1989 Dec;63(12):5386–5392. doi: 10.1128/jvi.63.12.5386-5392.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hellen C. U., Lee C. K., Wimmer E. Determinants of substrate recognition by poliovirus 2A proteinase. J Virol. 1992 Jun;66(6):3330–3338. doi: 10.1128/jvi.66.6.3330-3338.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hofer F., Gruenberger M., Kowalski H., Machat H., Huettinger M., Kuechler E., Blaas D. Members of the low density lipoprotein receptor family mediate cell entry of a minor-group common cold virus. Proc Natl Acad Sci U S A. 1994 Mar 1;91(5):1839–1842. doi: 10.1073/pnas.91.5.1839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kajigaya S., Arakawa H., Kuge S., Koi T., Imura N., Nomoto A. Isolation and characterization of defective-interfering particles of poliovirus Sabin 1 strain. Virology. 1985 Apr 30;142(2):307–316. doi: 10.1016/0042-6822(85)90339-3. [DOI] [PubMed] [Google Scholar]
  21. Kaplan G., Lubinski J., Dasgupta A., Racaniello V. R. In vitro synthesis of infectious poliovirus RNA. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8424–8428. doi: 10.1073/pnas.82.24.8424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kim Y. N., Makino S. Characterization of a murine coronavirus defective interfering RNA internal cis-acting replication signal. J Virol. 1995 Aug;69(8):4963–4971. doi: 10.1128/jvi.69.8.4963-4971.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kirkegaard K. Mutations in VP1 of poliovirus specifically affect both encapsidation and release of viral RNA. J Virol. 1990 Jan;64(1):195–206. doi: 10.1128/jvi.64.1.195-206.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kuge S., Saito I., Nomoto A. Primary structure of poliovirus defective-interfering particle genomes and possible generation mechanisms of the particles. J Mol Biol. 1986 Dec 5;192(3):473–487. doi: 10.1016/0022-2836(86)90270-6. [DOI] [PubMed] [Google Scholar]
  25. Lawson M. A., Semler B. L. Picornavirus protein processing--enzymes, substrates, and genetic regulation. Curr Top Microbiol Immunol. 1990;161:49–87. [PubMed] [Google Scholar]
  26. Lee W. M., Monroe S. S., Rueckert R. R. Role of maturation cleavage in infectivity of picornaviruses: activation of an infectosome. J Virol. 1993 Apr;67(4):2110–2122. doi: 10.1128/jvi.67.4.2110-2122.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Liljeström P., Lusa S., Huylebroeck D., Garoff H. In vitro mutagenesis of a full-length cDNA clone of Semliki Forest virus: the small 6,000-molecular-weight membrane protein modulates virus release. J Virol. 1991 Aug;65(8):4107–4113. doi: 10.1128/jvi.65.8.4107-4113.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mattion N. M., Reilly P. A., DiMichele S. J., Crowley J. C., Weeks-Levy C. Attenuated poliovirus strain as a live vector: expression of regions of rotavirus outer capsid protein VP7 by using recombinant Sabin 3 viruses. J Virol. 1994 Jun;68(6):3925–3933. doi: 10.1128/jvi.68.6.3925-3933.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mizutani S., Colonno R. J. In vitro synthesis of an infectious RNA from cDNA clones of human rhinovirus type 14. J Virol. 1985 Nov;56(2):628–632. doi: 10.1128/jvi.56.2.628-632.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Molla A., Jang S. K., Paul A. V., Reuer Q., Wimmer E. Cardioviral internal ribosomal entry site is functional in a genetically engineered dicistronic poliovirus. Nature. 1992 Mar 19;356(6366):255–257. doi: 10.1038/356255a0. [DOI] [PubMed] [Google Scholar]
  31. Molla A., Paul A. V., Schmid M., Jang S. K., Wimmer E. Studies on dicistronic polioviruses implicate viral proteinase 2Apro in RNA replication. Virology. 1993 Oct;196(2):739–747. doi: 10.1006/viro.1993.1531. [DOI] [PubMed] [Google Scholar]
  32. Novak J. E., Kirkegaard K. Coupling between genome translation and replication in an RNA virus. Genes Dev. 1994 Jul 15;8(14):1726–1737. doi: 10.1101/gad.8.14.1726. [DOI] [PubMed] [Google Scholar]
  33. Percy N., Barclay W. S., Sullivan M., Almond J. W. A poliovirus replicon containing the chloramphenicol acetyltransferase gene can be used to study the replication and encapsidation of poliovirus RNA. J Virol. 1992 Aug;66(8):5040–5046. doi: 10.1128/jvi.66.8.5040-5046.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Pfister T., Egger D., Bienz K. Poliovirus subviral particles associated with progeny RNA in the replication complex. J Gen Virol. 1995 Jan;76(Pt 1):63–71. doi: 10.1099/0022-1317-76-1-63. [DOI] [PubMed] [Google Scholar]
  35. Pilipenko E. V., Blinov V. M., Agol V. I. Gross rearrangements within the 5'-untranslated region of the picornaviral genomes. Nucleic Acids Res. 1990 Jun 11;18(11):3371–3375. doi: 10.1093/nar/18.11.3371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Pogue G. P., Hall T. C. The requirement for a 5' stem-loop structure in brome mosaic virus replication supports a new model for viral positive-strand RNA initiation. J Virol. 1992 Feb;66(2):674–684. doi: 10.1128/jvi.66.2.674-684.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Porter D. C., Ansardi D. C., Choi W. S., Morrow C. D. Encapsidation of genetically engineered poliovirus minireplicons which express human immunodeficiency virus type 1 Gag and Pol proteins upon infection. J Virol. 1993 Jul;67(7):3712–3719. doi: 10.1128/jvi.67.7.3712-3719.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Porter D. C., Ansardi D. C., Morrow C. D. Encapsidation of poliovirus replicons encoding the complete human immunodeficiency virus type 1 gag gene by using a complementation system which provides the P1 capsid protein in trans. J Virol. 1995 Mar;69(3):1548–1555. doi: 10.1128/jvi.69.3.1548-1555.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Quadt R., Ishikawa M., Janda M., Ahlquist P. Formation of brome mosaic virus RNA-dependent RNA polymerase in yeast requires coexpression of viral proteins and viral RNA. Proc Natl Acad Sci U S A. 1995 May 23;92(11):4892–4896. doi: 10.1073/pnas.92.11.4892. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Richards O. C., Ehrenfeld E. Poliovirus RNA replication. Curr Top Microbiol Immunol. 1990;161:89–119. doi: 10.1007/978-3-642-75602-3_4. [DOI] [PubMed] [Google Scholar]
  41. Rohll J. B., Percy N., Ley R., Evans D. J., Almond J. W., Barclay W. S. The 5'-untranslated regions of picornavirus RNAs contain independent functional domains essential for RNA replication and translation. J Virol. 1994 Jul;68(7):4384–4391. doi: 10.1128/jvi.68.7.4384-4391.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Rossmann M. G., Arnold E., Erickson J. W., Frankenberger E. A., Griffith J. P., Hecht H. J., Johnson J. E., Kamer G., Luo M., Mosser A. G. Structure of a human common cold virus and functional relationship to other picornaviruses. Nature. 1985 Sep 12;317(6033):145–153. doi: 10.1038/317145a0. [DOI] [PubMed] [Google Scholar]
  43. Rueckert R. R., Wimmer E. Systematic nomenclature of picornavirus proteins. J Virol. 1984 Jun;50(3):957–959. doi: 10.1128/jvi.50.3.957-959.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Sherry B., Rueckert R. Evidence for at least two dominant neutralization antigens on human rhinovirus 14. J Virol. 1985 Jan;53(1):137–143. doi: 10.1128/jvi.53.1.137-143.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Sommergruber W., Ahorn H., Zöphel A., Maurer-Fogy I., Fessl F., Schnorrenberg G., Liebig H. D., Blaas D., Kuechler E., Skern T. Cleavage specificity on synthetic peptide substrates of human rhinovirus 2 proteinase 2A. J Biol Chem. 1992 Nov 5;267(31):22639–22644. [PubMed] [Google Scholar]
  46. Wimmer E., Hellen C. U., Cao X. Genetics of poliovirus. Annu Rev Genet. 1993;27:353–436. doi: 10.1146/annurev.ge.27.120193.002033. [DOI] [PubMed] [Google Scholar]

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