Abstract
The development of effective purification protocols has permitted evaluation of the resistance of isolated scrapie prions to inactivation by UV irradiation at 254 nm. Prions were irradiated on ice with doses of UV light ranging up to 120,000 J/m2. UV dosimetry experiments, performed with Saccharomyces cerevisiae plasmid DNA or eucaryotic cells, indicated that under these experimental conditions an incident UV dose of 10 J/m2 formed 2 thymine dimers per 5.1 X 10(6) daltons of eucaryotic cell DNA. The D37 values for scrapie prions ranged from 17,000 to 22,000 J/m2; D37 values were also determined for virus, viroid, and enzyme controls. The number of pyrimidine dimers formed was correlated with the D37 values obtained for irradiated prions and target nucleic acids. The D37 value for bacteriophage M13, 6.5 J/m2, occurred at a dose that would form 0.56 dimers per target genome; the D37 for potato spindle tuber viroid, 4,800 J/m2, occurred at a dose that would form about 24 dimers per target viroid. The D37 value for an EcoRI restriction site, a target of 12 bases, occurred at a dose that would correspond to the formation of 0.89 thymine dimers per target site. The D37 value for prions occurred at a dose that would form 1 dimer in every 4 bases of single-stranded target nucleic acid. If the putative scrapie nucleic acid were double-stranded and readily repairable after UV damage, then the prion D37 value could reflect a nucleic acid molecule of 30 to 45 base pairs. While the D37 value for prions fell within the range of pure protein targets, our experiments cannot eliminate the possibility that a prion contains a small, highly protected nucleic acid molecule.
Full text
PDF







Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Alper T., Cramp W. A., Haig D. A., Clarke M. C. Does the agent of scrapie replicate without nucleic acid? Nature. 1967 May 20;214(5090):764–766. doi: 10.1038/214764a0. [DOI] [PubMed] [Google Scholar]
- Alper T., Haig D. A., Clarke M. C. The exceptionally small size of the scrapie agent. Biochem Biophys Res Commun. 1966 Feb 3;22(3):278–284. doi: 10.1016/0006-291x(66)90478-5. [DOI] [PubMed] [Google Scholar]
- Alper T., Haig D. A., Clarke M. C. The scrapie agent: evidence against its dependence for replication on intrinsic nucleic acid. J Gen Virol. 1978 Dec;41(3):503–516. doi: 10.1099/0022-1317-41-3-503. [DOI] [PubMed] [Google Scholar]
- Blinkhorn C., Jones M. N. The effect of surfactant structure on ribonuclease A-surfactant interactions. Biochem J. 1973 Nov;135(3):547–549. doi: 10.1042/bj1350547. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bolton D. C., McKinley M. P., Prusiner S. B. Identification of a protein that purifies with the scrapie prion. Science. 1982 Dec 24;218(4579):1309–1311. doi: 10.1126/science.6815801. [DOI] [PubMed] [Google Scholar]
- Bolton D. C., Meyer R. K., Prusiner S. B. Scrapie PrP 27-30 is a sialoglycoprotein. J Virol. 1985 Feb;53(2):596–606. doi: 10.1128/jvi.53.2.596-606.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Diener T. O., McKinley M. P., Prusiner S. B. Viroids and prions. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5220–5224. doi: 10.1073/pnas.79.17.5220. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Diener T. O., Schneider I. R., Smith D. R. Potato spindle tuber viroid. XI. A comparison of the ultraviolet light sensitivities of PSTV, tobacco ringspot virus, and its satellite. Virology. 1974 Feb;57(2):577–581. doi: 10.1016/0042-6822(74)90198-6. [DOI] [PubMed] [Google Scholar]
- GOLDE A., LATARJET R., VIGIER P. [Isotypical interference in vitro by Rous virus inactivated by ultraviolet rays]. C R Hebd Seances Acad Sci. 1961 Dec 4;253:2782–2784. [PubMed] [Google Scholar]
- Gajdusek D. C. Hypothesis: interference with axonal transport of neurofilament as a common pathogenetic mechanism in certain diseases of the central nervous system. N Engl J Med. 1985 Mar 14;312(11):714–719. doi: 10.1056/NEJM198503143121110. [DOI] [PubMed] [Google Scholar]
- Hines J. C., Ray D. S. Construction and characterization of new coliphage M13 cloning vectors. Gene. 1980 Nov;11(3-4):207–218. doi: 10.1016/0378-1119(80)90061-x. [DOI] [PubMed] [Google Scholar]
- LATARJET R., MORENNE P., BERGER R. Un appareil simple pour le dosage des rayonnements ultraviolets émis par les lampes germicides. Ann Inst Pasteur (Paris) 1953 Aug;85(2):175–184. [PubMed] [Google Scholar]
- Latarjet R., Cramer R., Montagnier L. Inactivation, by UV-, x-, and gamma-radiations, of the infecting and transforming capacities of polyoma virus. Virology. 1967 Sep;33(1):104–111. doi: 10.1016/0042-6822(67)90098-0. [DOI] [PubMed] [Google Scholar]
- Latarjet R., Muel B., Haig D. A., Clarke M. C., Alper T. Inactivation of the scrapie agent by near monochromatic ultraviolet light. Nature. 1970 Sep 26;227(5265):1341–1343. doi: 10.1038/2271341a0. [DOI] [PubMed] [Google Scholar]
- McKinley M. P., Bolton D. C., Prusiner S. B. A protease-resistant protein is a structural component of the scrapie prion. Cell. 1983 Nov;35(1):57–62. doi: 10.1016/0092-8674(83)90207-6. [DOI] [PubMed] [Google Scholar]
- McKinley M. P., Braunfeld M. B., Bellinger C. G., Prusiner S. B. Molecular characteristics of prion rods purified from scrapie-infected hamster brains. J Infect Dis. 1986 Jul;154(1):110–120. doi: 10.1093/infdis/154.1.110. [DOI] [PubMed] [Google Scholar]
- Merz P. A., Rohwer R. G., Kascsak R., Wisniewski H. M., Somerville R. A., Gibbs C. J., Jr, Gajdusek D. C. Infection-specific particle from the unconventional slow virus diseases. Science. 1984 Jul 27;225(4660):437–440. doi: 10.1126/science.6377496. [DOI] [PubMed] [Google Scholar]
- Oesch B., Westaway D., Wälchli M., McKinley M. P., Kent S. B., Aebersold R., Barry R. A., Tempst P., Teplow D. B., Hood L. E. A cellular gene encodes scrapie PrP 27-30 protein. Cell. 1985 Apr;40(4):735–746. doi: 10.1016/0092-8674(85)90333-2. [DOI] [PubMed] [Google Scholar]
- Prusiner S. B., Cochran S. P., Groth D. F., Downey D. E., Bowman K. A., Martinez H. M. Measurement of the scrapie agent using an incubation time interval assay. Ann Neurol. 1982 Apr;11(4):353–358. doi: 10.1002/ana.410110406. [DOI] [PubMed] [Google Scholar]
- Prusiner S. B., Groth D. F., Bolton D. C., Kent S. B., Hood L. E. Purification and structural studies of a major scrapie prion protein. Cell. 1984 Aug;38(1):127–134. doi: 10.1016/0092-8674(84)90533-6. [DOI] [PubMed] [Google Scholar]
- Prusiner S. B., Groth D. F., Cochran S. P., Masiarz F. R., McKinley M. P., Martinez H. M. Molecular properties, partial purification, and assay by incubation period measurements of the hamster scrapie agent. Biochemistry. 1980 Oct 14;19(21):4883–4891. doi: 10.1021/bi00562a028. [DOI] [PubMed] [Google Scholar]
- Prusiner S. B. Novel proteinaceous infectious particles cause scrapie. Science. 1982 Apr 9;216(4542):136–144. doi: 10.1126/science.6801762. [DOI] [PubMed] [Google Scholar]
- Prusiner S. B. Prions. Sci Am. 1984 Oct;251(4):50–59. doi: 10.1038/scientificamerican1084-50. [DOI] [PubMed] [Google Scholar]
- Rauth A. M. The Physical State of Viral Nucleic Acid and the Sensitivity of Viruses to Ultraviolet Light. Biophys J. 1965 May;5(3):257–273. doi: 10.1016/s0006-3495(65)86715-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
- Rohwer R. G. Scrapie infectious agent is virus-like in size and susceptibility to inactivation. Nature. 1984 Apr 12;308(5960):658–662. doi: 10.1038/308658a0. [DOI] [PubMed] [Google Scholar]