In vivo transfer of barley stripe mosaic hordeivirus ribonucleotides to the 5′ terminus of maize stripe tenuivirus RNAs

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The National Academy of Sciences

RESUMO

The Tenuivirus maize stripe virus (MStV) shares many properties with viruses in the genus Phlebovirus of the family Bunyaviridae. Besides genome organization and gene expression strategies, one property shared by these plant- and vertebrate-infecting viruses is that transcription gives rise to virus-specific mRNAs containing nonviral 5′-terminal nucleotide sequences. The 5′-terminal nucleotides are believed to be derived from host mRNA sequences as a result of “cap-snatching.” We investigated whether specific nucleotide sequences could serve as primer donors for cap-snatching in vivo. Barley (Hordeum vulgare) plants were singly and doubly infected with MStV and the Hordeivirus barley stripe mosaic virus (BSMV). A reverse transcription–PCR assay was used to identify chimeric BSMV/MStV RNAs. Specific reverse transcription–PCR products were detected from doubly infected plants by using one PCR primer corresponding to the 5′ termini of the BSMV RNAs (α, β, and γ) and a second primer complementary to MStV RNA 4. The resulting cDNAs were cloned, and nucleotide sequence analysis showed them to be chimeric, containing BSMV 5′-terminal sequences as well as MStV RNA 4 sequences. All clones contained the BSMV RNA 5′ primer nucleotide sequence, but they also showed characteristics common to Tenuivirus mRNAs. More than 80% of the clones contained BSMV RNA nucleotides not present on the PCR primer. Several lacked the exact 5′ terminus of MStV RNA 4, a feature also seen for viruses in the Bunyaviridae. These data show that heterologous virus RNAs (BSMV) can serve as primer donors for MStV mRNA capped RNA-primed transcription in doubly infected plants.

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