Nuclear ligation of RNA 5'-OH kinase products in tRNA.
Mouse L-cell nuclei incorporate gamma-32P from ATP in vitro predominantly in 5'-monophosphoryl termini and internal phosphodiester bonds with a nonrandom nearest-neighbor distribution. In the presence of 1 microgram of alpha-amanitin per ml the gamma-32P showed a time-dependent appearance in RNA bands which migrated with mature tRNA species but not with pre-tRNA and 5S RNA. The gamma-32P was found in internal phosphodiester bonds as shown by alkaline phosphatase resistance and was identified in 3'-monophosphates after RNase T2, T1, and A digestion. The specificity of this incorporation was indicated by a limited number of labeled oligonucleotides from a T1 digest and identification of 70 to 80% of the 32P label as Cp on complete digestion of the eluted tRNA band. We also observed transiently [gamma-32P]ATP-labeled RNA bands (in 5'-monophosphate positions) that were 32 to 45 nucleotides long. The results presented suggest splicing of several mouse L-cell tRNA species in isolated nuclei which involve the RNA 5'-OH kinase products as intermediates.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000494:pharmacology; D000546:Amanitins / Q000494:pharmacology; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D007739:L Cells; D051379:Mice; D010758:Phosphorus / Q000378:metabolism; D011116:Polynucleotide 5'-Hydroxyl-Kinase / Q000494:pharmacology; D012326:RNA Splicing; D012343:RNA, Transfer / Q000378:metabolism*; D012260:Ribonucleases / Q000494:pharmacology; D014544:Uridine Triphosphate / Q000378:metabolism
D000546:Amanitins; D010758:Phosphorus; D000255:Adenosine Triphosphate; D012343:RNA, Transfer; D011116:Polynucleotide 5'-Hydroxyl-Kinase; D012260:Ribonucleases; D000469:Alkaline Phosphatase; D014544:Uridine Triphosphate