Summary auto-generated
This 1989 study characterizes the genome organization of lettuce necrotic yellows virus (LNYV), a plant rhabdovirus. Researchers isolated the viral genomic RNA, estimated at approximately 13,000 nucleotides, and constructed complementary DNA (cDNA) clones. Northern blot analysis of infected Nicotiana glutinosa plants revealed five distinct polyadenylated viral mRNAs (cRNAs) ranging from 900 to 6,450 nucleotides in size. Individual recombinant cDNA clones hybridized to four of these cRNAs, which together represented about 95% of the viral genome sequence. Based on their sizes and coding capacities, the five cRNAs were provisionally assigned to encode the five structural proteins: the large L protein, envelope glycoprotein G, nucleocapsid protein N, non-structural NS protein, and matrix protein M. The researchers confirmed that cRNA 3 encodes the nucleocapsid protein by expressing a fusion protein from a cDNA clone that reacted specifically with a monoclonal antibody against the N protein. These findings represent the first detailed characterization of a plant rhabdovirus subgroup 1 genome and provide insights into LNYV gene expression and replication strategies.
Key findings
- LNYV genomic RNA consists of approximately 13,000 nucleotides of negative-sense, single-stranded RNA similar to other plant rhabdoviruses
- Five distinct polyadenylated viral mRNAs (cRNAs) were detected in infected plants, ranging from 900 to 6,450 nucleotides, representing about 95% of the viral genome
- cRNAs are transcribed from at least five LNYV genes encoding the major structural proteins: L, G, N, NS, and M proteins
- cRNA 3 (1,650 nt) was confirmed as the nucleocapsid protein mRNA through expression of a fusion protein recognized by anti-N protein antibodies
- Gene organization shows that genes for L and G proteins are adjacent, as are genes for N and NS proteins, consistent with other rhabdovirus genome arrangements
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Abstract
The negative-stranded genomic RNA of lettuce necrotic yellows virus (LNYV) was isolated and estimated by denaturing agarose gel electrophoresis to consist of about 13000 nucleotides (nt). Hybridization of randomly labelled LNYV RNA to poly(A)+ RNA from infected Nicotiana glutinosa plants revealed the presence of five distinct complementary RNA species (cRNAs) ranging in size from 900 to 6450 nt. Individual recombinant cDNA clones derived from LNYV RNA hybridized to four of these cRNA species. Each cRNA contained unique sequences which together represented about 95% of the viral genome. From their size range and sequence complexity, it is assumed that the cRNAs represent messenger RNAs encoding the five LNYV structural proteins. The mRNA for the nucleocapsid protein is cRNA 3 because a recombinant cDNA expression vector clone containing sequences which hybridize to cRNA 3 produced a polypeptide which was detected with a monoclonal antibody specific for the nucleocapsid protein.