Phosphoryl migration during the chemical synthesis of RNA.
By the use of high sensitivity assay systems, we have measured the occurrence of strand scission and phosphoryl migration that accompany the deblocking of chemically synthesized oligoribonucleotides. Substantial phosphoryl migration was observed for both enzymatically derived poly(uridylic acid) and synthetic uridine oligoribonucleotides 2'-O-protected with the 1-(2-fluorophenyl)-4-methoxypiperidin-4-yl (Fpmp) group, when these species were subjected to the acidic conditions suggested for Fpmp deprotection. Strand scission occurred in parallel and could be demonstrated readily by 5'-32P end labeling, but not by 3'-32P end labeling, of the acid-treated oligoribonucleotides. Increasing the pH of the deprotection solution and decreasing the temperature at which the deprotection was accomplished diminished both phosphoryl migration and strand scission. A mechanism that can rationalize these results is discussed.
D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009843:Oligoribonucleotides / Q000138:chemical synthesis; D010710:Phosphates / Q000737:chemistry*; D010880:Piperidines / Q000494:pharmacology; D011072:Poly U / Q000737:chemistry; D012313:RNA / Q000138:chemical synthesis* / Q000378:metabolism; D014529:Uridine / Q000737:chemistry
D009843:Oligoribonucleotides; D010710:Phosphates; D010880:Piperidines; D011072:Poly U; D012313:RNA; D014529:Uridine