A search for the first region in the genomic RNA of a potyvirus to be encapsidated during the assembly of virus particles in vivo has been undertaken. Protoplasts were collected at various times after inoculation and fragments of viral RNA that were protected from nuclease degradation were isolated from extracts and identified by RT-PCR procedures. Nuclease-resistant fragments of viral RNA were not detected in protoplasts that had been infected for 30 min. However, such fragments were present in protoplasts collected 40 min after inoculation and these contained a region at or near the 5' terminus of the viral RNA. Protoplasts that had been infected for 45 min or longer contained full-length viral RNA in a nuclease-resistant form. These results suggest that assembly of virus particles begins with the interaction of coat protein subunits with the 5' terminal region of progeny viral RNA molecules.


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