Functional significance of a hepta nucleotide motif present at the junction of Cucumber mosaic virus satellite RNA multimers in helper-virus dependent replication

Satellite RNAs (satRNA) associated with Cucumber mosaic virus (CMV) have been shown to generate multimers during replication. We have discovered that multimers of a CMV satRNA generated in the absence of its helper virus (HV) are characterized by the addition of a hepta nucleotide motif (HNM) at the monomer junctions. Here, we evaluated the functional significance of HNM in HV-dependent replication by ectopically expressing wild type and mutant forms of satRNA multimers in planta either in (+) or (−)-strand polarity. Comparative replication profiles revealed that (−)-strand multimers with complementary HNM (cHNM) are the preferred initial templates for HV-dependent replication than (−)-strand monomers and multimers lacking the cHNM. Further mutational analyses of the HNM accentuate that preservation of the sequence and native length of HNM is obligatory for efficient replication of satRNA. A model implicating the significance of HNM in HV-dependent production of monomeric and multimeric forms of satRNA is presented.

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► SatRNA mutilmers with hepta nucleotide motif (HNM) are produced in the absence of its helper virus.
► (−)-strand multimers with HNM are better templates than (−)-strand monomers.
► Preservation of the HNM sequence and length is obligatory for efficient replication.