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. 1989 Jul;63(7):2977–2986. doi: 10.1128/jvi.63.7.2977-2986.1989

Measles virus synthesizes both leaderless and leader-containing polyadenylated RNAs in vivo.

S J Castaneda 1, T C Wong 1
PMCID: PMC250852  PMID: 2470923

Abstract

The minus-sense RNA genome of measles virus serves as a template for synthesizing plus-sense RNAs of genomic length (antigenomes) and subgenomic length [poly(A)+ RNAs]. To elucidate how these different species are produced in vivo, RNA synthesized from the 3'-proximal N gene was characterized by Northern RNA blot and RNase protection analyses. The results showed that measles virus produced three size classes of plus-sense N-containing RNA species corresponding to monocistronic N RNA, bicistronic NP RNA, and antigenomes. Unlike vesicular stomatitis virus, measles virus does not produce a detectable free plus-sense leader RNA. Instead, although antigenomes invariably contain a leader sequence, monocistronic and bicistronic poly(A)+ N-containing RNAs are synthesized either without or with a leader sequence. We cloned and characterized a full-length cDNA representing a product of the latter type of synthesis. mRNAs and antigenomes appeared sequentially and in parallel with leaderless and leader-containing RNAs. These various RNA species accumulated concurrently throughout infection. However, cycloheximide preferentially inhibited accumulation of antigenomes and leader-containing RNA but not leaderless and subgenomic RNAs late in infection, suggesting that synthesis of the former RNA species requires a late protein function or a continuous supply of structural proteins or both. These results reveal a previously undescribed mechanism for RNA synthesis in measles virus.

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Selected References

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

  1. Abraham G., Banerjee A. K. Sequential transcription of the genes of vesicular stomatitis virus. Proc Natl Acad Sci U S A. 1976 May;73(5):1504–1508. doi: 10.1073/pnas.73.5.1504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alkhatib G., Briedis D. J. The predicted primary structure of the measles virus hemagglutinin. Virology. 1986 Apr 30;150(2):479–490. doi: 10.1016/0042-6822(86)90312-0. [DOI] [PubMed] [Google Scholar]
  3. Arnheiter H., Davis N. L., Wertz G., Schubert M., Lazzarini R. A. Role of the nucleocapsid protein in regulating vesicular stomatitis virus RNA synthesis. Cell. 1985 May;41(1):259–267. doi: 10.1016/0092-8674(85)90079-0. [DOI] [PubMed] [Google Scholar]
  4. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Baker S. C., Moyer S. A. Encapsidation of Sendai virus genome RNAs by purified NP protein during in vitro replication. J Virol. 1988 Mar;62(3):834–838. doi: 10.1128/jvi.62.3.834-838.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ball L. A., White C. N. Order of transcription of genes of vesicular stomatitis virus. Proc Natl Acad Sci U S A. 1976 Feb;73(2):442–446. doi: 10.1073/pnas.73.2.442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Banerjee A. D., Abraham G., Colonno R. J. Vesicular stomatitis virus: mode of transcription. J Gen Virol. 1977 Jan;34(1):1–8. doi: 10.1099/0022-1317-34-1-1. [DOI] [PubMed] [Google Scholar]
  8. Banerjee A. K. Transcription and replication of rhabdoviruses. Microbiol Rev. 1987 Mar;51(1):66–87. doi: 10.1128/mr.51.1.66-87.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Bellini W. J., Englund G., Richardson C. D., Rozenblatt S., Lazzarini R. A. Matrix genes of measles virus and canine distemper virus: cloning, nucleotide sequences, and deduced amino acid sequences. J Virol. 1986 May;58(2):408–416. doi: 10.1128/jvi.58.2.408-416.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Bellini W. J., Englund G., Rozenblatt S., Arnheiter H., Richardson C. D. Measles virus P gene codes for two proteins. J Virol. 1985 Mar;53(3):908–919. doi: 10.1128/jvi.53.3.908-919.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Billeter M. A., Baczko K., Schmid A., Ter Meulen V. Cloning of DNA corresponding to four different measles virus genomic regions. Virology. 1984 Jan 15;132(1):147–159. doi: 10.1016/0042-6822(84)90099-0. [DOI] [PubMed] [Google Scholar]
  12. Blumberg B. M., Crowley J. C., Silverman J. I., Menonna J., Cook S. D., Dowling P. C. Measles virus L protein evidences elements of ancestral RNA polymerase. Virology. 1988 Jun;164(2):487–497. doi: 10.1016/0042-6822(88)90563-6. [DOI] [PubMed] [Google Scholar]
  13. Blumberg B. M., Kolakofsky D. Intracellular vesicular stomatitis virus leader RNAs are found in nucleocapsid structures. J Virol. 1981 Nov;40(2):568–576. doi: 10.1128/jvi.40.2.568-576.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Blumberg B. M., Leppert M., Kolakofsky D. Interaction of VSV leader RNA and nucleocapsid protein may control VSV genome replication. Cell. 1981 Mar;23(3):837–845. doi: 10.1016/0092-8674(81)90448-7. [DOI] [PubMed] [Google Scholar]
  15. Carroll A. R., Wagner R. R. Adenosine-5'-O-(3-thiotriphosphate) as an affinity probe for studying leader RNA's transcribed by vesicular stomatitis virus. J Biol Chem. 1979 Oct 10;254(19):9339–9341. [PubMed] [Google Scholar]
  16. Casey J., Davidson N. Rates of formation and thermal stabilities of RNA:DNA and DNA:DNA duplexes at high concentrations of formamide. Nucleic Acids Res. 1977;4(5):1539–1552. doi: 10.1093/nar/4.5.1539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Cattaneo R., Kaelin K., Baczko K., Billeter M. A. Measles virus editing provides an additional cysteine-rich protein. Cell. 1989 Mar 10;56(5):759–764. doi: 10.1016/0092-8674(89)90679-x. [DOI] [PubMed] [Google Scholar]
  18. Chambers P., Millar N. S., Bingham R. W., Emmerson P. T. Molecular cloning of complementary DNA to Newcastle disease virus, and nucleotide sequence analysis of the junction between the genes encoding the haemagglutinin-neuraminidase and the large protein. J Gen Virol. 1986 Mar;67(Pt 3):475–486. doi: 10.1099/0022-1317-67-3-475. [DOI] [PubMed] [Google Scholar]
  19. Chanda P. K., Banerjee A. K. Identification of promoter-proximal oligonucleotides and a unique dinucleotide, pppGpC, from in vitro transcription products of vesicular stomatitis virus. J Virol. 1981 Jul;39(1):93–103. doi: 10.1128/jvi.39.1.93-103.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  21. Collins P. L., Hightower L. E., Ball L. A. Transcriptional map for Newcastle disease virus. J Virol. 1980 Sep;35(3):682–693. doi: 10.1128/jvi.35.3.682-693.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Collins P. L., Wertz G. W., Ball L. A., Hightower L. E. Coding assignments of the five smaller mRNAs of Newcastle disease virus. J Virol. 1982 Sep;43(3):1024–1031. doi: 10.1128/jvi.43.3.1024-1031.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Colonno R. J., Banerjee A. K. A unique RNA species involved in initiation of vesicular stomatitis virus RNA transcription in vitro. Cell. 1976 Jun;8(2):197–204. doi: 10.1016/0092-8674(76)90003-9. [DOI] [PubMed] [Google Scholar]
  24. Colonno R. J., Banerjee A. K. Mapping and initiation studies on the leader RNA of vesicular stomatitis virus. Virology. 1977 Mar;77(1):260–268. doi: 10.1016/0042-6822(77)90423-8. [DOI] [PubMed] [Google Scholar]
  25. Crowley J. C., Dowling P. C., Menonna J., Silverman J. I., Schuback D., Cook S. D., Blumberg B. M. Sequence variability and function of measles virus 3' and 5' ends and intercistronic regions. Virology. 1988 Jun;164(2):498–506. doi: 10.1016/0042-6822(88)90564-8. [DOI] [PubMed] [Google Scholar]
  26. Davis N. L., Wertz G. W. Synthesis of vesicular stomatitis virus negative-strand RNA in vitro: dependence on viral protein synthesis. J Virol. 1982 Mar;41(3):821–832. doi: 10.1128/jvi.41.3.821-832.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Dowling P. C., Blumberg B. M., Menonna J., Adamus J. E., Cook P., Crowley J. C., Kolakofsky D., Cook S. D. Transcriptional map of the measles virus genome. J Gen Virol. 1986 Sep;67(Pt 9):1987–1992. doi: 10.1099/0022-1317-67-9-1987. [DOI] [PubMed] [Google Scholar]
  28. Emerson S. U. Reconstitution studies detect a single polymerase entry site on the vesicular stomatitis virus genome. Cell. 1982 Dec;31(3 Pt 2):635–642. doi: 10.1016/0092-8674(82)90319-1. [DOI] [PubMed] [Google Scholar]
  29. Flamand A., Delagneau J. F. Transcriptional mapping of rabies virus in vivo. J Virol. 1978 Nov;28(2):518–523. doi: 10.1128/jvi.28.2.518-523.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Fyrberg E. A., Kindle K. L., Davidson N., Kindle K. L. The actin genes of Drosophila: a dispersed multigene family. Cell. 1980 Feb;19(2):365–378. doi: 10.1016/0092-8674(80)90511-5. [DOI] [PubMed] [Google Scholar]
  31. Glazier K., Raghow R., Kingsbury D. W. Regulation of Sendai virus transcription: evidence for a single promoter in vivo. J Virol. 1977 Mar;21(3):863–871. doi: 10.1128/jvi.21.3.863-871.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Gupta K. C., Kingsbury D. W. Polytranscripts of Sendai virus do not contain intervening polyadenylate sequences. Virology. 1985 Feb;141(1):102–109. doi: 10.1016/0042-6822(85)90186-2. [DOI] [PubMed] [Google Scholar]
  33. Herman R. C., Lazzarini R. A. Vesicular stomatitis virus RNA polymerase can read through the boundary between the leader and N genes in vitro. J Virol. 1981 May;38(2):792–796. doi: 10.1128/jvi.38.2.792-796.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Herman R. C., Schubert M., Keene J. D., Lazzarini R. A. Polycistronic vesicular stomatitis virus RNA transcripts. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4662–4665. doi: 10.1073/pnas.77.8.4662. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Hill V. M., Summers D. F. Synthesis of VSV RNPs in vitro by cellular VSV RNPs added to uninfected HeLa cell extracts: VSV protein requirements for replication in vitro. Virology. 1982 Dec;123(2):407–419. doi: 10.1016/0042-6822(82)90273-2. [DOI] [PubMed] [Google Scholar]
  36. Iverson L. E., Rose J. K. Sequential synthesis of 5'-proximal vesicular stomatitis virus mRNA sequences. J Virol. 1982 Oct;44(1):356–365. doi: 10.1128/jvi.44.1.356-365.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Kurilla M. G., Piwnica-Worms H., Keene J. D. Rapid and transient localization of the leader RNA of vesicular stomatitis virus in the nuclei of infected cells. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5240–5244. doi: 10.1073/pnas.79.17.5240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Kurilla M. G., Stone H. O., Keene J. D. RNA sequence and transcriptional properties of the 3' end of the Newcastle disease virus genome. Virology. 1985 Sep;145(2):203–212. doi: 10.1016/0042-6822(85)90154-0. [DOI] [PubMed] [Google Scholar]
  39. Lazzarini R. A., Keene J. D., Schubert M. The origins of defective interfering particles of the negative-strand RNA viruses. Cell. 1981 Oct;26(2 Pt 2):145–154. doi: 10.1016/0092-8674(81)90298-1. [DOI] [PubMed] [Google Scholar]
  40. Leppert M., Rittenhouse L., Perrault J., Summers D. F., Kolakofsky D. Plus and minus strand leader RNAs in negative strand virus-infected cells. Cell. 1979 Nov;18(3):735–747. doi: 10.1016/0092-8674(79)90127-2. [DOI] [PubMed] [Google Scholar]
  41. MARMUR J., DOTY P. Determination of the base composition of deoxyribonucleic acid from its thermal denaturation temperature. J Mol Biol. 1962 Jul;5:109–118. doi: 10.1016/s0022-2836(62)80066-7. [DOI] [PubMed] [Google Scholar]
  42. Masters P. S., Samuel C. E. Detection of in vivo synthesis of polycistronic mRNAs of vesicular stomatitis virus. Virology. 1984 Apr 30;134(2):277–286. doi: 10.1016/0042-6822(84)90297-6. [DOI] [PubMed] [Google Scholar]
  43. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Okayama H., Berg P. A cDNA cloning vector that permits expression of cDNA inserts in mammalian cells. Mol Cell Biol. 1983 Feb;3(2):280–289. doi: 10.1128/mcb.3.2.280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Patton J. T., Davis N. L., Wertz G. W. N protein alone satisfies the requirement for protein synthesis during RNA replication of vesicular stomatitis virus. J Virol. 1984 Feb;49(2):303–309. doi: 10.1128/jvi.49.2.303-309.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Perrault J., Clinton G. M., McClure M. A. RNP template of vesicular stomatitis virus regulates transcription and replication functions. Cell. 1983 Nov;35(1):175–185. doi: 10.1016/0092-8674(83)90220-9. [DOI] [PubMed] [Google Scholar]
  47. Piwnica-Worms H., Keene J. D. Sequential synthesis of small capped RNA transcripts in vitro by vesicular stomatitis virus. Virology. 1983 Feb;125(1):206–218. doi: 10.1016/0042-6822(83)90074-0. [DOI] [PubMed] [Google Scholar]
  48. Richardson C., Hull D., Greer P., Hasel K., Berkovich A., Englund G., Bellini W., Rima B., Lazzarini R. The nucleotide sequence of the mRNA encoding the fusion protein of measles virus (Edmonston strain): a comparison of fusion proteins from several different paramyxoviruses. Virology. 1986 Dec;155(2):508–523. doi: 10.1016/0042-6822(86)90212-6. [DOI] [PubMed] [Google Scholar]
  49. Rima B. K., Baczko K., Clarke D. K., Curran M. D., Martin S. J., Billeter M. A., ter Meulen V. Characterization of clones for the sixth (L) gene and a transcriptional map for morbilliviruses. J Gen Virol. 1986 Sep;67(Pt 9):1971–1978. doi: 10.1099/0022-1317-67-9-1971. [DOI] [PubMed] [Google Scholar]
  50. Rozenblatt S., Eizenberg O., Ben-Levy R., Lavie V., Bellini W. J. Sequence homology within the morbilliviruses. J Virol. 1985 Feb;53(2):684–690. doi: 10.1128/jvi.53.2.684-690.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. 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]
  52. Seifried A. S., Albrecht P., Milstien J. B. Characterization of an RNA-dependent RNA polymerase activity associated with measles virus. J Virol. 1978 Mar;25(3):781–787. doi: 10.1128/jvi.25.3.781-787.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Shioda T., Hidaka Y., Kanda T., Shibuta H., Nomoto A., Iwasaki K. Sequence of 3,687 nucleotides from the 3' end of Sendai virus genome RNA and the predicted amino acid sequences of viral NP, P and C proteins. Nucleic Acids Res. 1983 Nov 11;11(21):7317–7330. doi: 10.1093/nar/11.21.7317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Shioda T., Iwasaki K., Shibuta H. Determination of the complete nucleotide sequence of the Sendai virus genome RNA and the predicted amino acid sequences of the F, HN and L proteins. Nucleic Acids Res. 1986 Feb 25;14(4):1545–1563. doi: 10.1093/nar/14.4.1545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Sprague J., Condra J. H., Arnheiter H., Lazzarini R. A. Expression of a recombinant DNA gene coding for the vesicular stomatitis virus nucleocapsid protein. J Virol. 1983 Feb;45(2):773–781. doi: 10.1128/jvi.45.2.773-781.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Testa D., Chanda P. K., Banerjee A. K. Unique mode of transcription in vitro by Vesicular stomatitis virus. Cell. 1980 Aug;21(1):267–275. doi: 10.1016/0092-8674(80)90134-8. [DOI] [PubMed] [Google Scholar]
  57. Udem S. A. Measles virus: conditions for the propagation and purification of infectious virus in high yield. J Virol Methods. 1984 Feb;8(1-2):123–136. doi: 10.1016/0166-0934(84)90046-6. [DOI] [PubMed] [Google Scholar]
  58. Wechsler S. L., Fields B. N. Intracellular synthesis of measles virus-specified polypeptides. J Virol. 1978 Jan;25(1):285–297. doi: 10.1128/jvi.25.1.285-297.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Wertz G. W., Levine M. RNA synthesis by vesicular stomatitis virus and a small plaque mutant: effects of cycloheximide. J Virol. 1973 Aug;12(2):253–264. doi: 10.1128/jvi.12.2.253-264.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Wilde A., Morrison T. Structural and functional characterization of Newcastle disease virus polycistronic RNA species. J Virol. 1984 Jul;51(1):71–76. doi: 10.1128/jvi.51.1.71-76.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Wong T. C., Hirano A. Functional cDNA library for efficient expression of measles virus-specific gene products in primate cells. J Virol. 1986 Jan;57(1):343–348. doi: 10.1128/jvi.57.1.343-348.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Yoshikawa Y., Mizumoto K., Yamanouchi K. Characterization of messenger RNAs of measles virus. J Gen Virol. 1986 Dec;67(Pt 12):2807–2812. doi: 10.1099/0022-1317-67-12-2807. [DOI] [PubMed] [Google Scholar]

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