Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1990 Feb;64(2):774–783. doi: 10.1128/jvi.64.2.774-783.1990

cis elements and trans-acting factors involved in dimer formation of murine leukemia virus RNA.

A C Prats 1, C Roy 1, P A Wang 1, M Erard 1, V Housset 1, C Gabus 1, C Paoletti 1, J L Darlix 1
PMCID: PMC249172  PMID: 2153242

Abstract

The genetic material of all retroviruses examined so far consists of two identical RNA molecules joined at their 5' ends by the dimer linkage structure (DLS). Since the precise location of the DLS as well as the mechanism and role(s) of RNA dimerization remain unclear, we analyzed the dimerization process of Moloney murine leukemia virus (MoMuLV) genomic RNA. For this purpose we derived an in vitro model for RNA dimerization. By using this model, murine leukemia virus RNA was shown to form dimeric molecules. Deletion mutagenesis in the 620-nucleotide leader of MoMuLV RNA showed that the dimer promoting sequences are located within the encapsidation element Psi between positions 215 and 420. Furthermore, hybridization assays in which DNA oligomers were used to probe monomer and dimer forms of MoMuLV RNA indicated that the DLS probably maps between positions 280 and 330 from the RNA 5' end. Also, retroviral nucleocapsid protein was shown to catalyze dimerization of MoMuLV RNA and to be tightly bound to genomic dimer RNA in virions. These results suggest that MoMuLV RNA dimerization and encapsidation are probably controlled by the same cis element, Psi, and trans-acting factor, nucleocapsid protein, and thus might be linked during virion formation.

Full text

PDF
777

Images in this article

777Image
on p.777
778Image
on p.778
779Image
on p.779
780Image
on p.780
781Image
on p.781

Selected References

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

  1. Bader J. P., Ray D. A. Configurational variants of oncornavirus RNAs. J Virol. 1976 Sep;19(3):810–819. doi: 10.1128/jvi.19.3.810-819.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bender W., Chien Y. H., Chattopadhyay S., Vogt P. K., Gardner M. B., Davidson N. High-molecular-weight RNAs of AKR, NZB, and wild mouse viruses and avian reticuloendotheliosis virus all have similar dimer structures. J Virol. 1978 Mar;25(3):888–896. doi: 10.1128/jvi.25.3.888-896.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bender W., Davidson N. Mapping of poly(A) sequences in the electron microscope reveals unusual structure of type C oncornavirus RNA molecules. Cell. 1976 Apr;7(4):595–607. doi: 10.1016/0092-8674(76)90210-5. [DOI] [PubMed] [Google Scholar]
  4. Canaani E., Helm K. V., Duesberg P. Evidence for 30-40S RNA as precursor of the 60-70S RNA of Rous sarcoma virus. Proc Natl Acad Sci U S A. 1973 Feb;70(2):401–405. doi: 10.1073/pnas.70.2.401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Darlix J. L. Control of Rous sarcoma virus RNA translation and packaging by the 5' and 3' untranslated sequences. J Mol Biol. 1986 Jun 5;189(3):421–434. doi: 10.1016/0022-2836(86)90314-1. [DOI] [PubMed] [Google Scholar]
  6. Darlix J. L., Spahr P. F., Bromley P. A. Analysis of Rous sarcoma virus (RSV) RNA structure by means of specific nucleases. Virology. 1978 Oct 15;90(2):317–329. doi: 10.1016/0042-6822(78)90316-1. [DOI] [PubMed] [Google Scholar]
  7. Darlix J. L., Spahr P. F., Bromley P. A., Jaton J. C. In vitro, the major ribosome binding site on Rous sarcoma virus RNA does not contain the nucleotide sequence coding for the N-terminal amino acids of the gag gene product. J Virol. 1979 Feb;29(2):597–611. doi: 10.1128/jvi.29.2.597-611.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fan H., Baltimore D. RNA metabolism of murine leukemia virus: detection of virus-specific RNA sequences in infected and uninfected cells and identification of virus-specific messenger RNA. J Mol Biol. 1973 Oct 15;80(1):93–117. doi: 10.1016/0022-2836(73)90235-0. [DOI] [PubMed] [Google Scholar]
  9. Fan H., Jaenisch R., MacIsaac P. Low-multiplicity infection of Moloney murine leukemia virus in mouse cells: effect on number of viral DNA copies and virus production in producer cells. J Virol. 1978 Dec;28(3):802–809. doi: 10.1128/jvi.28.3.802-809.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ganoza M. C., Kofoid E. C., Marlière P., Louis B. G. Potential secondary structure at translation-initiation sites. Nucleic Acids Res. 1987 Jan 12;15(1):345–360. doi: 10.1093/nar/15.1.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gorelick R. J., Henderson L. E., Hanser J. P., Rein A. Point mutants of Moloney murine leukemia virus that fail to package viral RNA: evidence for specific RNA recognition by a "zinc finger-like" protein sequence. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8420–8424. doi: 10.1073/pnas.85.22.8420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Havron A., Sperling J. Specificity of photochemical cross-linking in protein-nucleic acid complexes: identification of the interacting residues in RNase- pyrimidine nucleotide complex. Biochemistry. 1977 Dec 13;16(25):5631–5635. doi: 10.1021/bi00644a038. [DOI] [PubMed] [Google Scholar]
  13. Herr W. Nucleotide sequence of AKV murine leukemia virus. J Virol. 1984 Feb;49(2):471–478. doi: 10.1128/jvi.49.2.471-478.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kung H. J., Hu S., Bender W., Bailey J. M., Davidson N., Nicolson M. O., McAllister R. M. RD-114, baboon, and woolly monkey viral RNA's compared in size and structure. Cell. 1976 Apr;7(4):609–620. doi: 10.1016/0092-8674(76)90211-7. [DOI] [PubMed] [Google Scholar]
  15. Leis J., Baltimore D., Bishop J. M., Coffin J., Fleissner E., Goff S. P., Oroszlan S., Robinson H., Skalka A. M., Temin H. M. Standardized and simplified nomenclature for proteins common to all retroviruses. J Virol. 1988 May;62(5):1808–1809. doi: 10.1128/jvi.62.5.1808-1809.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Maisel J., Bender W., Hu S., Duesberg P. H., Davidson N. Structure of 50 to 70S RNA from Moloney sarcoma viruses. J Virol. 1978 Jan;25(1):384–394. doi: 10.1128/jvi.25.1.384-394.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mangel W. F., Delius H., Duesberg P. H. Structure and molecular weight of the 60-70S RNA and the 30-40S RNA of the Rous sarcoma virus. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4541–4545. doi: 10.1073/pnas.71.11.4541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mann R., Baltimore D. Varying the position of a retrovirus packaging sequence results in the encapsidation of both unspliced and spliced RNAs. J Virol. 1985 May;54(2):401–407. doi: 10.1128/jvi.54.2.401-407.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mann R., Mulligan R. C., Baltimore D. Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus. Cell. 1983 May;33(1):153–159. doi: 10.1016/0092-8674(83)90344-6. [DOI] [PubMed] [Google Scholar]
  20. Merregaert J., Janowski M., Reddy E. P. Nucleotide sequence of a radiation leukemia virus genome. Virology. 1987 May;158(1):88–102. doi: 10.1016/0042-6822(87)90241-8. [DOI] [PubMed] [Google Scholar]
  21. Miller A. D., Verma I. M. Two base changes restore infectivity to a noninfectious molecular clone of Moloney murine leukemia virus (pMLV-1). J Virol. 1984 Jan;49(1):214–222. doi: 10.1128/jvi.49.1.214-222.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Moelling K., Sykora K. W., Dittmar K. E., Scott A., Watson K. F. The isolation of avian viral RNA and polypeptides. J Biol Chem. 1979 May 25;254(10):3738–3742. [PubMed] [Google Scholar]
  23. Munroe S. H., Duthie R. S. Splice site consensus sequences are preferentially accessible to nucleases in isolated adenovirus RNA. Nucleic Acids Res. 1986 Nov 11;14(21):8447–8465. doi: 10.1093/nar/14.21.8447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Murti K. G., Bondurant M., Tereba A. Secondary structural features in the 70S RNAs of Moloney murine leukemia and Rous sarcoma viruses as observed by electron microscopy. J Virol. 1981 Jan;37(1):411–419. doi: 10.1128/jvi.37.1.411-419.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Méric C., Darlix J. L., Spahr P. F. It is Rous sarcoma virus protein P12 and not P19 that binds tightly to Rous sarcoma virus RNA. J Mol Biol. 1984 Mar 15;173(4):531–538. doi: 10.1016/0022-2836(84)90396-6. [DOI] [PubMed] [Google Scholar]
  26. Méric C., Goff S. P. Characterization of Moloney murine leukemia virus mutants with single-amino-acid substitutions in the Cys-His box of the nucleocapsid protein. J Virol. 1989 Apr;63(4):1558–1568. doi: 10.1128/jvi.63.4.1558-1568.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Panganiban A. T., Fiore D. Ordered interstrand and intrastrand DNA transfer during reverse transcription. Science. 1988 Aug 26;241(4869):1064–1069. doi: 10.1126/science.2457948. [DOI] [PubMed] [Google Scholar]
  28. Prats A. C., De Billy G., Wang P., Darlix J. L. CUG initiation codon used for the synthesis of a cell surface antigen coded by the murine leukemia virus. J Mol Biol. 1989 Jan 20;205(2):363–372. doi: 10.1016/0022-2836(89)90347-1. [DOI] [PubMed] [Google Scholar]
  29. Prats A. C., Sarih L., Gabus C., Litvak S., Keith G., Darlix J. L. Small finger protein of avian and murine retroviruses has nucleic acid annealing activity and positions the replication primer tRNA onto genomic RNA. EMBO J. 1988 Jun;7(6):1777–1783. doi: 10.1002/j.1460-2075.1988.tb03008.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schulein M., Burnette W. N., August J. T. Stoichiometry and specificity of binding of Rauscher oncovirus 10,000-dalton (p10) structural protein to nucleic acids. J Virol. 1978 Apr;26(1):54–60. doi: 10.1128/jvi.26.1.54-60.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schwartzberg P., Colicelli J., Goff S. P. Deletion mutants of Moloney murine leukemia virus which lack glycosylated gag protein are replication competent. J Virol. 1983 May;46(2):538–546. doi: 10.1128/jvi.46.2.538-546.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sen A., Todaro G. J. The genome-associated, specific RNA binding proteins of avian and mammalian type C viruses. Cell. 1977 Jan;10(1):91–99. doi: 10.1016/0092-8674(77)90143-x. [DOI] [PubMed] [Google Scholar]
  33. Shinnick T. M., Lerner R. A., Sutcliffe J. G. Nucleotide sequence of Moloney murine leukaemia virus. Nature. 1981 Oct 15;293(5833):543–548. doi: 10.1038/293543a0. [DOI] [PubMed] [Google Scholar]
  34. Stoltzfus C. M., Snyder P. N. Structure of B77 sarcoma virus RNA: stabilization of RNA after packaging. J Virol. 1975 Nov;16(5):1161–1170. doi: 10.1128/jvi.16.5.1161-1170.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tukalo M. A., Kubler M. D., Kern D., Mougel M., Ehresmann C., Ebel J. P., Ehresmann B., Giegé R. trans-Diamminedichloroplatinum(II), a reversible RNA-protein cross-linking agent. Application to the ribosome and to an aminoacyl-tRNA synthetase/tRNA complex. Biochemistry. 1987 Aug 11;26(16):5200–5208. doi: 10.1021/bi00390a045. [DOI] [PubMed] [Google Scholar]
  36. Varani G., Wimberly B., Tinoco I., Jr Conformation and dynamics of an RNA internal loop. Biochemistry. 1989 Sep 19;28(19):7760–7772. doi: 10.1021/bi00445a036. [DOI] [PubMed] [Google Scholar]
  37. Vollenweider H. J., Sogo J. M., Koller T. A routine method for protein-free spreading of double- and single-stranded nucleic acid molecules. Proc Natl Acad Sci U S A. 1975 Jan;72(1):83–87. doi: 10.1073/pnas.72.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Zuker M., Stiegler P. Optimal computer folding of large RNA sequences using thermodynamics and auxiliary information. Nucleic Acids Res. 1981 Jan 10;9(1):133–148. doi: 10.1093/nar/9.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES