Study of TMV assembly with heterologous RNA containing the origin-of-assembly sequence

The assembly of tobacco mosaic virus (TMV) is initiated by a specific reaction between a capsid protein oligomer and an origin-of-assembly region (OAS) located 900 nucleotides from the 3' terminus of virion RNA. Packaging is then completed by rod elongation both in the 5' and 3' directions. The temporal order of the direction of elongation and the characteristics of the reaction were studied by analysis of the in vitro assembly reaction between strain U1 protein oligomers and transcripts containing a strain U1 OAS embedded at different positions in heterologous RNA. The results confirm that elongation in the 5' direction starts very soon after the initiation reaction and is completed rapidly, within minutes. Packaging in the 3' direction is slower and does not appear to commence until 5' rod formation is complete. The reaction of strain U2 protein with the strain U1 OAS initiates rapidly, but elongation occurs only in the 5' direction; 3' packaging does not occur except when the OAS is at or near the 5' terminus, in which case elongation in the 3' direction initiates without delay with either the U1 or U2 protein. Pauses occur during elongation in the 3' direction at an average of 320 nucleotides, indicating a packaging periodicity of about six to eight helical turns.