Evidence that DNA is present in abnormal tubulofilamentous structures found in scrapie.
Abnormal tubulofilamentous structures have been identified in electron micrographs of thin sections and negatively stained impression grids prepared from brains of animals with scrapie and other spongiform encephalopathies, and we showed that such tubules contain a core of filamentous structures resembling scrapie-associated fibrils (SAF). We treated impression grids from brains of scrapie-infected hamsters with several substances that bind to or cleave proteins and nucleic acids to see if they had any effect on the abnormal tubulofilamentous structures. Treatment with three proteolytic enzymes reduced the caliber of the tubules from about 50 nm to 30 nm; subsequent treatment of the 30-nm tubules with DNase I left many typical SAF as well as transitional forms in which twisted SAF emerged from tubules. DNase treatment of the original thicker tubules had no effect, and no SAF were seen on grids. Treatment of the 30-nm tubules with any of three other nucleases (micrococcal, mung bean, and BAL-31) also produced SAF. However, treatment with RNase A had no effect either on the original 50-nm tubules or on the 30-nm tubules produced by proteolysis. Detergent treatment of any of the preparations produced SAF. Treatment with ethidium bromide resulted in staining of the tubules that was inhibited by magnesium ions. The data suggest that the abnormal tubulofilamentous particles found in spongiform encephalopathies may consist of an outer cylinder of protein, an inner cylinder of DNA, and an innermost core of SAF.
D000818:Animals; D001921:Brain / Q000382:microbiology* / Q000473:pathology / Q000648:ultrastructure; D006224:Cricetinae; D004279:DNA, Viral / Q000302:isolation & purification*; D003851:Deoxyribonucleases; D005260:Female; D008647:Mesocricetus; D008854:Microscopy, Electron; D011328:Prions / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D012608:Scrapie / Q000473:pathology*
D004279:DNA, Viral; D011328:Prions; D003851:Deoxyribonucleases