Lucerne transient streak sobemovirus (LTSV) supports the replication of a small, 322-nucleotide, untranslated virusoid (vLTSV) that has an extensively base-paired, viroid-like structure. Since vLTSV does not code for its own proteins or share sequence homology with its helper virus (LTSV), it is presumed that it uses structural motifs to signal the helper virus (and host) machinery for its replication. In order to elucidate these structural domains, insertion-deletion mutations were introduced to disrupt the secondary structure. Infectivity assays of these mutants showed that they were all lethal, except a 9-nucleotide, palindromic insertion, which preserved the overall rod-like structure of the virusoid. Sequence analysis of cDNA clones prepared from progeny virusoid RNA revealed that the palindromic sequence was replicated up to twelve days of infection but discarded afterwards. Results indicate that vLTSV has an optimum size and secondary structure for replication and packaging within the LTSV helper virus.

Lucerne transient streak sobemovirus (LTSV) supports the replication of a small, 322-nucleotide, untranslated virusoid (vLTSV) that has an extensively base-paired, viroid-like structure. Since vLTSV does not code for its own proteins or share sequence homology with its helper virus (LTSV), it is presumed that it uses structural motifs to signal the helper virus (and host) machinery for its replication. In order to elucidate these structural domains, insertion-deletion mutations were introduced to disrupt the secondary structure. Infectivity assays of these mutants showed that they were all lethal, except a 9-nucleotide, palindromic insertion, which preserved the overall rod-like structure of the virusoid. Sequence analysis of cDNA clones prepared from progeny virusoid RNA revealed that the palindromic sequence was replicated up to twelve days of infection but discarded afterwards. Results indicate that vLTSV has an optimum size and secondary structure for replication and packaging within the LTSV helper virus.