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. 1996 Aug;70(8):5100–5105. doi: 10.1128/jvi.70.8.5100-5105.1996

Accumulation of alfalfa mosaic virus RNAs 1 and 2 requires the encoded proteins in cis.

C M van Rossum 1, M L Garcia 1, J F Bol 1
PMCID: PMC190464  PMID: 8764017

Abstract

RNAs 1 and 2 of the tripartite genome of alfalfa mosaic virus (A1MV) encode the replicase proteins P1 and P2, respectively. P1 expressed in transgenic plants (P1 plants) can be used in trans to support replication of A1MV RNAs 2 and 3, and P2 expressed in transgenic plants (P2 plants) can be used in trans to support replication of A1MV RNAs 1 and 3. Wild-type RNA 1 was able to coreplicate with RNAs 2 and 3 in P1 plants, but this ability was abolished by frameshifts or deletions in the P1 gene of RNA 1. Similarly, wild-type RNA 2 coreplicated with RNAs 1 and 3 in P2 plants, but frameshifts or deletions in the P2 gene of RNA 2 interfered with this replication. Apparently, the P1 and P2 genes are required in cis for the accumulation of RNAs 1 and 2, respectively. Point mutations in the GDD motif of the P2 gene in RNA 2 interfered with accumulation of RNA 2 in P2 plants, indicating that replication of RNA 2 is linked to its translation into a functional protein. Plants transformed with both the P1 and P2 genes (P12 plants) accumulate replicase activity that is able to replicate RNA 3 in trans. An analysis of the time course of the accumulation of RNAs 1, 2, and 3 in protoplasts of P12 plants supported the conclusion that translation and replication are tightly coupled for A1MV RNAs 1 and 2 but not for RNA 3.

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Selected References

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  1. Bol J. F., van Vloten-Doting L., Jaspars E. M. A functional equivalence of top component a RNA and coat protein in the initiation of infection by alfalfa mosaic virus. Virology. 1971 Oct;46(1):73–85. doi: 10.1016/0042-6822(71)90007-9. [DOI] [PubMed] [Google Scholar]
  2. Brederode F. T., Taschner P. E., Posthumus E., Bol J. F. Replicase-mediated resistance to alfalfa mosaic virus. Virology. 1995 Mar 10;207(2):467–474. doi: 10.1006/viro.1995.1106. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. De Graaff M., Coscoy L., Jaspars E. M. Localization and biochemical characterization of alfalfa mosaic virus replication complexes. Virology. 1993 Jun;194(2):878–881. doi: 10.1006/viro.1993.1335. [DOI] [PubMed] [Google Scholar]
  5. De Graaff M., Man in't Veld M. R., Jaspars E. M. In vitro evidence that the coat protein of alfalfa mosaic virus plays a direct role in the regulation of plus and minus RNA synthesis: implications for the life cycle of alfalfa mosaic virus. Virology. 1995 Apr 20;208(2):583–589. doi: 10.1006/viro.1995.1189. [DOI] [PubMed] [Google Scholar]
  6. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Houwing C. J., Jaspars E. M. Coat protein stimulates replication complexes of alfalfa mosaic virus to produce virion RNAs in vitro. Biochimie. 1993;75(7):617–621. doi: 10.1016/0300-9084(93)90068-4. [DOI] [PubMed] [Google Scholar]
  9. Koonin E. V. The phylogeny of RNA-dependent RNA polymerases of positive-strand RNA viruses. J Gen Virol. 1991 Sep;72(Pt 9):2197–2206. doi: 10.1099/0022-1317-72-9-2197. [DOI] [PubMed] [Google Scholar]
  10. Loesch-Fries L. S., Halk E. L., Nelson S. E., Krahn K. J. Human leukocyte interferon does not inhibit alfalfa mosaic virus in protoplasts or tobacco tissue. Virology. 1985 Jun;143(2):626–629. doi: 10.1016/0042-6822(85)90402-7. [DOI] [PubMed] [Google Scholar]
  11. Neeleman L., Van der Vossen E. A., Bol J. F. Infection of tobacco with alfalfa mosaic virus cDNAs sheds light on the early function of the coat protein. Virology. 1993 Oct;196(2):883–887. doi: 10.1006/viro.1993.1551. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Quadt R., Rosdorff H. J., Hunt T. W., Jaspars E. M. Analysis of the protein composition of alfalfa mosaic virus RNA-dependent RNA polymerase. Virology. 1991 May;182(1):309–315. doi: 10.1016/0042-6822(91)90674-z. [DOI] [PubMed] [Google Scholar]
  14. Taschner P. E., van Marle G., Brederode F. T., Tumer N. E., Bol J. F. Plants transformed with a mutant alfalfa mosaic virus coat protein gene are resistant to the mutant but not to wild-type virus. Virology. 1994 Sep;203(2):269–276. doi: 10.1006/viro.1994.1484. [DOI] [PubMed] [Google Scholar]
  15. Taschner P. E., van der Kuyl A. C., Neeleman L., Bol J. F. Replication of an incomplete alfalfa mosaic virus genome in plants transformed with viral replicase genes. Virology. 1991 Apr;181(2):445–450. doi: 10.1016/0042-6822(91)90876-d. [DOI] [PubMed] [Google Scholar]
  16. Van Bokhoven H., Le Gall O., Kasteel D., Verver J., Wellink J., Van Kammen A. B. Cis- and trans-acting elements in cowpea mosaic virus RNA replication. Virology. 1993 Aug;195(2):377–386. doi: 10.1006/viro.1993.1387. [DOI] [PubMed] [Google Scholar]
  17. Van der Kuyl A. C., Neeleman L., Bol J. F. Role of alfalfa mosaic virus coat protein in regulation of the balance between viral plus and minus strand RNA synthesis. Virology. 1991 Nov;185(1):496–499. doi: 10.1016/0042-6822(91)90807-n. [DOI] [PubMed] [Google Scholar]
  18. Weiland J. J., Dreher T. W. Cis-preferential replication of the turnip yellow mosaic virus RNA genome. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6095–6099. doi: 10.1073/pnas.90.13.6095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. White K. A., Bancroft J. B., Mackie G. A. Coding capacity determines in vivo accumulation of a defective RNA of clover yellow mosaic virus. J Virol. 1992 May;66(5):3069–3076. doi: 10.1128/jvi.66.5.3069-3076.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. de Groot R. J., van der Most R. G., Spaan W. J. The fitness of defective interfering murine coronavirus DI-a and its derivatives is decreased by nonsense and frameshift mutations. J Virol. 1992 Oct;66(10):5898–5905. doi: 10.1128/jvi.66.10.5898-5905.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. van Dun C. M., van Vloten-Doting L., Bol J. F. Expression of alfalfa mosaic virus cDNA1 and 2 in transgenic tobacco plants. Virology. 1988 Apr;163(2):572–578. doi: 10.1016/0042-6822(88)90298-x. [DOI] [PubMed] [Google Scholar]
  22. van der Kuyl A. C., Neeleman L., Bol J. F. Complementation and recombination between alfalfa mosaic virus RNA3 mutants in tobacco plants. Virology. 1991 Aug;183(2):731–738. doi: 10.1016/0042-6822(91)91002-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. van der Kuyl A. C., Neeleman L., Bol J. F. Deletion analysis of cis- and trans-acting elements involved in replication of alfalfa mosaic virus RNA 3 in vivo. Virology. 1991 Aug;183(2):687–694. doi: 10.1016/0042-6822(91)90997-p. [DOI] [PubMed] [Google Scholar]
  24. van der Vossen E. A., Neeleman L., Bol J. F. Early and late functions of alfalfa mosaic virus coat protein can be mutated separately. Virology. 1994 Aug 1;202(2):891–903. doi: 10.1006/viro.1994.1411. [DOI] [PubMed] [Google Scholar]
  25. van der Vossen E. A., Neeleman L., Bol J. F. Role of the 5' leader sequence of alfalfa mosaic virus RNA 3 in replication and translation of the viral RNA. Nucleic Acids Res. 1993 Mar 25;21(6):1361–1367. doi: 10.1093/nar/21.6.1361. [DOI] [PMC free article] [PubMed] [Google Scholar]

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