Mapping of the RD phenotype of the Nancy strain of coxsackievirus B3.
The RD variants of group B coxsackieviruses differ from their parental strains in having the ability to replicate in a human rhabdomyosarcoma cell line, RD. The nucleotide sequence of the P1 region of the RD variant of coxsackievirus B3 strain Nancy (CB3NRD) was determined by sequencing cloned cDNAs, obtained by PCR amplification. A comparison between the established nucleotide sequence and that of the P1 region from the parental virus revealed 12 point mutations which corresponded to six amino acid replacements. To identify if the P1 region is responsible for the phenotype of CB3NRD, a chimeric virus was constructed, using an infectious cDNA clone of CB3. The P1 region of the infectious cDNA was replaced by cDNA fragments from CB3N (parental strain Nancy) or CB3NRD and the resulting recombinants were assayed for their ability to infect and replicate in RD cells. The results showed that the RD phenotype of CB3NRD maps in the P1 region. Furthermore, a chimera which only contained the 5' part of the P1 region derived from CB3NRD and the remaining P1 sequence from CB3N was able to replicate in RD cells, suggesting that the VP2 polypeptide contains at least one determinant for the RD phenotype.
Lindberg
A M
AM
Department of Medical Genetics, Uppsala University, Sweden.
Crowell
R L
RL
Zell
R
R
Kandolf
R
R
Pettersson
U
U
D001483:Base Sequence; D002213:Capsid / Q000235:genetics; D036022:Capsid Proteins; D002460:Cell Line; D002678:Chimera; D029822:Enterovirus B, Human / Q000235:genetics* / Q000254:growth & development; D014644:Genetic Variation; D016679:Genome, Viral; D008969:Molecular Sequence Data; D009710:Nucleotide Mapping / Q000379:methods; D010641:Phenotype; D012208:Rhabdomyosarcoma / Q000382:microbiology*; D014407:Tumor Cells, Cultured; D014779:Virus Replication
D036022:Capsid Proteins; C442554:VP2 protein, Echovirus