In-vitro translation of cucumoviral satellites. III. Translational efficiencies of cucumber mosaic virus-associated RNA 5 sequence variants can be related to the predicted secondary structures of their first 55 nucleotides.
The cucumber mosaic virus (CMV) satellites D- and S-CARNA 5 (CARNA 5 = Cucumber mosaic virus-Associated RNA 5), their full-length cDNA clone transcripts, and DNA clone transcripts of their open reading frames (ORFs) were used as mRNAs in the wheat-germ in-vitro translation system. Natural D-CARNA 5 yielded an anomalously large polypeptide, while transcripts made from cDNA clones of D-CARNA 5 or its first ORF had no mRNA activity. Transcripts made from the second major ORF in D-CARNA 5 yielded a smaller product, consistent with its size. Natural S-CARNA 5 and its cDNA clone transcripts both yielded the two polypeptides previously reported, while transcripts of its only major ORF yielded exclusively the smaller of the two products. The potential for an alternate initiation codon, 36 nucleotides upstream, being the source of the larger of the two polypeptides was tested. The differences in the translational properties of D- and S-CARNA 5 were related to the predicted secondary structures of the first 55 nucleotides in these CARNA 5 sequence variants. The calculated free energies of the predicted hairpins correlated inversely with their in-vitro translational activities.
Microbiology and Plant Pathology Laboratory, A.R.S., U.S.D.A., Beltsville, MD 20705.
M T
MT
Steen
J M
JM
Kaper
C W
CW
Pleij
J N
JN
Hansen
D001483:Base Sequence; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000032:analysis*; D004926:Escherichia coli / Q000235:genetics; D014644:Genetic Variation; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000235:genetics*; D009690:Nucleic Acid Conformation; D014176:Protein Biosynthesis; D012313:RNA / Q000032:analysis*; D019244:RNA, Satellite; D012367:RNA, Viral / Q000032:analysis*
D004279:DNA, Viral; D019244:RNA, Satellite; D012367:RNA, Viral; D012313:RNA