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Defective interfering particles of poliovirus: mapping of the deletion and evidence that the deletions in the genomes of DI(1), (2) and (3) are located in the same region



Defective interfering particles of poliovirus: mapping of the deletion and evidence that the deletions in the genomes of DI(1), (2) and (3) are located in the same region



Journal of Molecular Biology 128(2): 179-196



The deletions in RNA of 3 defective interfering (DI) particles of poliovirus type 1 were located and their approximate extent determined by 3 methods. Digestion with [Escherichia coli] RNase III, of DI RNA yields the same 3'-terminal fragments as digestion with RNase III of standard virus RNA. The longest 3'-terminal fragment has a MW of 1.55 .times. 106. The deletions are probably located in the 5'-terminal half of the polio genome. Fingerprints of RNase T1-resistant oligonucleotides of all 3 DI RNA are identical and lack 4 large oligonucleotides as compared to the fingerprints of standard virus, suggesting that the deletions in all 3 DI RNA are located in the same region of the viral genome. The deletion-specific oligonucleotides were also within the 5'-terminal half of the viral genome by alkali fragmentation of the RNA and fingerprinting poly(A)-linked (3'-terminal) fragments of decreasing size. Virion RNA of DI(2) particles was annealed with denatured double-stranded RNA (RF) of standard virus and the hybrid heteroduplex molecules examined by EM. A single loop, about 900 nucleotides long and 20% from 1 end of the molecules, was observed. Both the size and extent of individual deletions is somewhat variable in different heteroduplex molecules, an observation suggesting heterogeneity in the size of the deletion in RNA of the DI(2) population. The DI RNA of poliovirus contain an internal that region of the viral genome known to specify the capsid polypeptides. This result provides an explanation as to why poliovirus DI particles are unable to synthesize viral coat proteins.

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Accession: 068524522

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PMID: 219204

DOI: 10.1016/0022-2836(79)90125-6


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