Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1986 Dec 22;14(24):9877–9896. doi: 10.1093/nar/14.24.9877

Terminal structure of hypovirulence-associated dsRNAs in the chestnut blight fungus Endothia parasitica.

S Hiremath, B L'Hostis, S A Ghabrial, R E Rhoads
PMCID: PMC341341  PMID: 3808956

Abstract

The 3'- and 5'-terminal sequences of the five large double-stranded RNA species (L-dsRNA; 4.5-6.0 X 10(6) daltons) of EP713, a hypovirulent strain of Endothia parasitica, were determined by mobility-shift and enzymatic methods. All the L-dsRNAs appeared to have identical terminal sequences. A heteropolymer sequence was found at one 3'-terminus and a poly(A) sequence of variable length at the other. It was possible to label only one 5'-terminus using polynucleotide kinase and [gamma-32P]ATP, and this was shown to be a poly(U) sequence of variable length. We propose that the dsRNAs have the following structure, where X represents a blocking group: (Formula: see text). A recombinant plasmid containing dsRNA-related sequences was constructed. Hybridization analysis using the recombinant probe indicated that the sequence homology among the L-dsRNAs extended beyond these terminal regions and was also shared by small dsRNAs (0.3-0.45 X 10(6) daltons).

Full text

PDF
9880

Images in this article

9881Image
on p.9881
9882Image
on p.9882
9884Image
on p.9884
9885Image
on p.9885
9886Image
on p.9886
9888Image
on p.9888
9890Image
on p.9890

Selected References

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

  1. Alwine J. C., Kemp D. J., Stark G. R. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5350–5354. doi: 10.1073/pnas.74.12.5350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anagnostakis S. L. Biological control of chestnut blight. Science. 1982 Jan 29;215(4532):466–471. doi: 10.1126/science.215.4532.466. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bruce A. G., Uhlenbeck O. C. Reactions at the termini of tRNA with T4 RNA ligase. Nucleic Acids Res. 1978 Oct;5(10):3665–3677. doi: 10.1093/nar/5.10.3665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cashdollar L. W., Esparza J., Hudson G. R., Chmelo R., Lee P. W., Joklik W. K. Cloning the double-stranded RNA genes of reovirus: sequence of the cloned S2 gene. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7644–7648. doi: 10.1073/pnas.79.24.7644. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Craig E. A., McCarthy B. J. Four Drosophila heat shock genes at 67B: characterization of recombinant plasmids. Nucleic Acids Res. 1980 Oct 10;8(19):4441–4457. doi: 10.1093/nar/8.19.4441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Diaz-Ruiz J. R., Kaper J. M. Cucumber mosaic virus-associated RNA 5. III. Little nucleotide sequence homology between CARNA 5 and helper RNA. Virology. 1977 Jul 1;80(1):204–213. doi: 10.1016/0042-6822(77)90393-2. [DOI] [PubMed] [Google Scholar]
  9. Donis-Keller H., Maxam A. M., Gilbert W. Mapping adenines, guanines, and pyrimidines in RNA. Nucleic Acids Res. 1977 Aug;4(8):2527–2538. doi: 10.1093/nar/4.8.2527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hannig E. M., Williams T. L., Leibowitz M. J. The internal polyadenylate tract of yeast killer virus M1 double-stranded RNA is variable in length. Virology. 1986 Jul 15;152(1):149–158. doi: 10.1016/0042-6822(86)90380-6. [DOI] [PubMed] [Google Scholar]
  12. Lockard R. E., Alzner-Deweerd B., Heckman J. E., MacGee J., Tabor M. W., RajBhandary U. L. Sequence analysis of 5'[32P] labeled mRNA and tRNA using polyacrylamide gel electrophoresis. Nucleic Acids Res. 1978 Jan;5(1):37–56. doi: 10.1093/nar/5.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Malek L. T., Eschenfeldt W. H., Munns T. W., Rhoads R. E. Heterogeneity of the 5' terminus of hen ovalbumin messenger ribonucleic acid. Nucleic Acids Res. 1981 Apr 10;9(7):1657–1673. doi: 10.1093/nar/9.7.1657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  16. Mossop D. W., Francki R. I. Association of RNA 3 with aphid transmission of cucumber mosaic virus. Virology. 1977 Aug;81(1):177–181. doi: 10.1016/0042-6822(77)90071-x. [DOI] [PubMed] [Google Scholar]
  17. Nuss D. L., Summers D. Variant dsRNAs associated with transmission-defective isolates of wound tumor virus represent terminally conserved remnants of genome segments. Virology. 1984 Mar;133(2):276–288. doi: 10.1016/0042-6822(84)90395-7. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Schneider I. R., White R. M. Tobacco ringspot virus codes for the coat protein of its satellite. Virology. 1976 Mar;70(1):244–246. doi: 10.1016/0042-6822(76)90264-6. [DOI] [PubMed] [Google Scholar]
  20. Shoulder A., Darby G., Minson T. RNA-RNA hybridisation using 125I-labelled RNA from tobacco necrosis virus and its satellite. Nature. 1974 Oct 25;251(5477):733–735. doi: 10.1038/251733a0. [DOI] [PubMed] [Google Scholar]
  21. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  22. Szekeres M., Brownstein B. H., Revel H. R., Haselkorn R. Terminal sequences of the bacteriophage phi 6 segmented dsRNA genome and its messenger RNAs. Virology. 1985 Apr 15;142(1):1–11. doi: 10.1016/0042-6822(85)90417-9. [DOI] [PubMed] [Google Scholar]
  23. Tsiapalis C. M., Dorson J. W., De Sante D. M., Bollum F. J. Terminal riboadenylate transferase: a polyadenylate polymerase from calf thymus gland. Biochem Biophys Res Commun. 1973 Feb 5;50(3):737–743. doi: 10.1016/0006-291x(73)91306-5. [DOI] [PubMed] [Google Scholar]
  24. Vary C. P., Vournakis J. N. RNase H-catalyzed site-specific deadenylylation of rabbit alpha- and beta- globin mRNAs. Secondary structure of 3'-noncoding regions. J Biol Chem. 1984 Mar 10;259(5):3299–3307. [PubMed] [Google Scholar]
  25. de Wachter R., Fiers W. Preparative two-dimensional polyacrylamide gel electrophoresis of 32 P-labeled RNA. Anal Biochem. 1972 Sep;49(1):184–197. doi: 10.1016/0003-2697(72)90257-6. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES