The effects of S-adenosyl methionine (AdoMet) and its analogues on the control of transcription and translation in vitro of the mRNA products of two cytoplasmic polyhedrosis viruses.
S-Adenosyl methionine (AdoMet) and several structurally related compounds were added to in vitro systems for the synthesis of single-stranded RNA by cytoplasmic polyhedrosis virus (CPV) types 1 and 2. The effects of these compounds were examined on the level of transcription and methylation of the RNA products. Of the compounds tested, five increased the polymerase activity in both viruses, the most effective being the D- and L-stereoisomers of S-adenosyl homocysteine (AdoHcy), and the least effective, adenosine. L-AdoHcy, unlike D-AdoHcy, was also a competitive inhibitor of RNA methylation in the presence of [3H]AdoMet. The different response of both viruses to D- and L-AdoHcy suggests that CPV virions contain at least two functionally distinct sites to which AdoMet, or its analogues, bind. One of these is the transcription control site, while the other is the active site(s) for RNA methylation. CPV RNA synthesised in the presence of the methyl donor AdoMet was more efficiently translated in vitro in a wheat-germ translation system than RNA synthesised in the presence of methylation inhibitors. Type 2 CPV-RNA transcripts had a greater degree of methylation than type 1 CPV transcripts and were more effective in stimulating protein synthesis in the translation system. It seems likely that the allosteric control of CPV polymerase by AdoMet and its analogues, and the methylation of the transcripts, ensures the effective transcription and translation of the CPV genome and the stability of the viral messenger RNA.
D000241:Adenosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D002474:Cell-Free System; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D005001:Ethionine / Q000031:analogs & derivatives* / Q000494:pharmacology; D006710:Homocysteine / Q000031:analogs & derivatives*; D007304:Insect Viruses / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D007700:Kinetics; D010944:Plants / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects*; D012333:RNA, Messenger / Q000235:genetics*; D012265:Ribonucleotides / Q000494:pharmacology; D012435:S-Adenosylhomocysteine / Q000494:pharmacology*; D012436:S-Adenosylmethionine / Q000494:pharmacology*; D013045:Species Specificity; D014158:Transcription, Genetic / Q000187:drug effects*; D014908:Triticum / Q000378:metabolism
D012333:RNA, Messenger; D012265:Ribonucleotides; D006710:Homocysteine; C012167:S-adenosylethionine; D012436:S-Adenosylmethionine; D012435:S-Adenosylhomocysteine; D012321:DNA-Directed RNA Polymerases; D000241:Adenosine; D005001:Ethionine