Nucleic Acids Research, 1994, Vol. 22, No. 6 1101-1107
© 1994
MOLECULAR BIOLOGY |
Endogenous viral complexes with long RNA cosediment with the agent of Creutzfeldt-Jakob Disease
Yale Medical School 310 Cedar Street, New Haven, CT 06510, USA
*To whom correspondence should be addressed
Received August 30, 1993. Revised January 28, 1994. Accepted January 28, 1994.
A class of viruslike agents that Induces Creutzfeldt-Jakob Disease (CJD) and scraple remains undefined at the molecular level. Several investigators believe this infectious agent Is constituted by a single host protein or ‘prion’, and have emphasized data that would seem to exclude the presence of any viral nucleic acids. However, more rigorous evaluations in scraple have shown reasonably abundant nucleic acids. Additionally, in highly purified 120S CJD preparations that have been treated with nucleases, RNAs as long as 6,000 bases have been detected. Few nucleic acids have been characterized in either scraple or CJD, but previous cloning experiments delineated relatively short LTR regions of the endogenous IAP retrovlrus In 120S CJD preparations. We therefore used specific primers encompassing the entire IAP genome to test for the presence of long viral RNAs, and here show 
5,000 contiguous bases of the IAP RNA genome can be recovered from reasonable amounts of starting brain. The 3' env region of IAP is comparably truncated in CJD and normal preparations, and we find no evidence for IAP transduction of CJD-speciflc sequences. Because IAP cores can coencapsidate unrelated sequences, and are unusually resistant to physical and chemical treatments, it was relevant to find if cosedimentlng cognate proteins of the IAP core, such as gag, could be detected. The predicted 65kd acidic gag protein, showing appropriate antlgenlc and nucleic acid binding features, was apparent in both one and 2-D Western blots. This data strongly indicates specific viral complexes cofractionate with the CJD agent. Interestingly, these nuclease resistant lAPs do not appear to be In morphologically recognizable ‘R’ particles. This cosedimentlng viral assembly therefore provides a paradigm for non-particulate CJD complexes in infectious preparations. In developing strategies to identify a CJD specific sequence, cosedimentlng lAPs can be used to assess the quality, length and recovery of RNAs extracted from highly resistant viral complexes.
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