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. 1988 May 11;16(9):4009–4023. doi: 10.1093/nar/16.9.4009

Autolytic processing of a phosphorothioate diester bond.

J M Buzayan 1, P A Feldstein 1, C Segrelles 1, G Bruening 1
PMCID: PMC336571  PMID: 2453843

Abstract

A small satellite RNA of tobacco ringspot virus replicates in tissues infected with tobacco ringspot virus and accumulates in virus capsids, forming virus-like particles. Previous research showed that multimeric forms of this satellite RNA have tandem repeats of the "monomeric" satellite RNA sequence of 359 or 360 nucleotide residues. The multimeric RNAs undergo autolytic processing at a specific CpA phosphodiester bond, the junction, to generate the monomeric RNA. We substituted phosphorothioate diester bonds for various sets of phosphodiester bonds, in dimeric and truncated forms of the satellite RNA. The degree of reduction in autolytic cleavage varied both with the sites of substitution and the size of the RNA molecules. Analyses of a product of the autolysis reaction suggest that one phosphorothioate diester bond most strongly interferes with processing, the one introduced at the CpA junction during its synthesis from adenosine-5'-0-(1-thiotriphosphate). However, extensive introduction of phosphorothioate diester bonds elsewhere in the molecule also decreased processing, possibly by altering conformation.

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Selected References

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

  1. Buzayan J. M., Gerlach W. L., Bruening G. Satellite tobacco ringspot virus RNA: A subset of the RNA sequence is sufficient for autolytic processing. Proc Natl Acad Sci U S A. 1986 Dec;83(23):8859–8862. doi: 10.1073/pnas.83.23.8859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Buzayan J. M., McNinch J. S., Schneider I. R., Bruening G. A nucleotide sequence rearrangement distinguishes two isolates of satellite tobacco ringspot virus RNA. Virology. 1987 Sep;160(1):95–99. doi: 10.1016/0042-6822(87)90049-3. [DOI] [PubMed] [Google Scholar]
  3. Davanloo P., Rosenberg A. H., Dunn J. J., Studier F. W. Cloning and expression of the gene for bacteriophage T7 RNA polymerase. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2035–2039. doi: 10.1073/pnas.81.7.2035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Eckstein F. Nucleoside phosphorothioates. Annu Rev Biochem. 1985;54:367–402. doi: 10.1146/annurev.bi.54.070185.002055. [DOI] [PubMed] [Google Scholar]
  5. Eckstein F., Romaniuk P. J., Connolly B. A. Stereochemistry of enzymic phosphoryl and nucleotidyl transfer. Methods Enzymol. 1982;87:197–212. doi: 10.1016/s0076-6879(82)87015-8. [DOI] [PubMed] [Google Scholar]
  6. Francki R. I. Plant virus satellites. Annu Rev Microbiol. 1985;39:151–174. doi: 10.1146/annurev.mi.39.100185.001055. [DOI] [PubMed] [Google Scholar]
  7. Frey P. A., Richard J. P., Ho H. T., Brody R. S., Sammons R. D., Sheu K. F. Stereochemistry of selected phosphotransferases and nucleotidyltransferases. Methods Enzymol. 1982;87:213–235. doi: 10.1016/s0076-6879(82)87016-x. [DOI] [PubMed] [Google Scholar]
  8. Haseloff J., Symons R. H. Chrysanthemum stunt viroid: primary sequence and secondary structure. Nucleic Acids Res. 1981 Jun 25;9(12):2741–2752. doi: 10.1093/nar/9.12.2741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kaper J. M., Tousignant M. E. Viral satellites: parasitic nucleic acids capable of modulating disease expression. Endeavour. 1984;8(4):194–200. doi: 10.1016/0160-9327(84)90084-x. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Prody G. A., Bakos J. T., Buzayan J. M., Schneider I. R., Bruening G. Autolytic processing of dimeric plant virus satellite RNA. Science. 1986 Mar 28;231(4745):1577–1580. doi: 10.1126/science.231.4745.1577. [DOI] [PubMed] [Google Scholar]
  12. Westheimer F. H. Why nature chose phosphates. Science. 1987 Mar 6;235(4793):1173–1178. doi: 10.1126/science.2434996. [DOI] [PubMed] [Google Scholar]

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