[Molecular genetic analysis of poliovirus infection]

Molecular genetic studies of the antigenicity and the attenuation phenotype of type 1 poliovirus were described. Antigenic sites were identified on the genome of type 1 poliovirus by the determination of nucleotide sequence of the genome of variants that were not neutralized by the neutralizing monoclonal antibodies. The solution of the crystal structure of poliovirus revealed that all mutations found as above are located at the surface of the virion and cluster into three distinct sites. These regions probably represent distinct antibody binding sites. To study expression of the attenuation phenotype of type 1 poliovirus, a number of recombinant polioviruses were constructed in vitro by using infectious complementary deoxyribonucleic acid clones of the virulent Mahoney and attenuated Sabin 1 strains of type 1 poliovirus. Biological tests including a monkey neurovirulence test were performed on the recombinants. The results indicated that the 5' noncoding region harbors a relatively strong determinant influencing the attenuation. Further studies revealed that an adenine residue (Mahoney type) at nucleotide position 480 importantly contribute to the expression of the neurovirulence phenotype. However, a guanine residue (Sabin 1 type) at position 480 was not sufficient for full expression of the attenuation phenotype encoded by this genome region. These results suggested that the expression of the attenuation phenotype depends on the highly ordered structure formed in the 5' noncoding sequence and that the formation of such a structure is possibly influenced by the nucleotide position 480. To investigate the structure and function of the 5' noncoding region, many insertion and deletion sequences were introduced into the genome region. Replication processes of the mutants were analysed and second-site mutations in the genome of the variants that partially restored the phenotypes of the parental viruses were identified. The results indicated that interactions between different loci, for example at around positions 200 and 500, are important for maintaining the viral replication efficiency.