Two autolytic processing reactions of a satellite RNA proceed with inversion of configuration.
Both polarities of the satellite RNA of tobacco ringspot virus occur in infected cells in multimeric forms which are capable of autolytic processing, using different sequences and structures [Feldstein, P.A., et al., Proc. Nat. Acad. Sci. USA (1990) 87 (in press)]. These transesterification reactions generate a 2',3'-cyclophosphate and a 5'-hydroxyl as the two new end groups. Cleavage is at a CpA for the (+) polarity RNA and at an ApG for the (-) polarity RNA. We enzymically synthesized oligoribonucleotides with processing capability and with specific 35S-labeled phosphorothioate diesters in the Rp configuration. After processing had occurred, the terminal nucleoside-2',3'-cyclophosphorothioate diester residues were recovered from the appropriate product by digestion with nuclease and phosphatase. Comparisons with specially prepared endo- and exoisomer reference compounds by thin layer chromatography and autoradiography revealed that the [35S]cytidine- and [35S]adenosine-2',3'-cyclophosphorothioate both were endo-isomers. The results are consistent with transesterification occurring by an inline SN2(P) attack of the 2'-hydroxyl group in the autolytic processing reactions of both polarities of the satellite RNA.
Department of Plant Pathology, College of Agricultural and Environmental Sciences, University of California, Davis 95616.
H
H
van Tol
J M
JM
Buzayan
P A
PA
Feldstein
F
F
Eckstein
G
G
Bruening
D001483:Base Sequence; D055598:Chemical Phenomena; D002621:Chemistry; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009690:Nucleic Acid Conformation; D009843:Oligoribonucleotides / Q000378:metabolism; D010942:Plant Viruses / Q000235:genetics*; D010947:Plants, Toxic; D012313:RNA / Q000235:genetics / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional; D019244:RNA, Satellite; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D014026:Nicotiana
D009843:Oligoribonucleotides; D019244:RNA, Satellite; D012367:RNA, Viral; D012313:RNA