3'-immature tRNA(Trp) is required for ribosome inactivation by gelonin,a plant RNA N-glycosidase.
Inactivation of ribosomes by gelonin, a ribosome-inactivating protein with RNA N-glycosidase activity on 28 S rRNA, requires macromolecular cofactors present in post-ribosomal supernatants. One of these cofactors has been purified from a rat liver extract and identified as an RNA about 70 nt long which in sequence analysis shows a high level of similarity with mammalian (bovine) tRNA(Trp). The pattern of the sequencing gel is consistent with the co-existence in the preparation of two 3'-immature tRNA(Trp) species, missing only A75, or both A75 and C74. In the presence of ATP, CTP and tRNA nucleotidyltransferase, the gelonin-stimulating RNA is a good acceptor of tryptophan. An oligodeoxynucleotide complementary to positions 55 to 72 of mammalian (bovine) tRNA(Trp) hybridizes with the gelonin-stimulating RNA as demonstrated by gel mobility shift and ribonuclease H digestion. The oligodeoxynucleotide-directed ribonuclease H treatment also abolishes the gelonin-promoting activity of crude preparations of RNA, giving strong evidence that the only active RNA is a tRNA(Trp)-like molecule.
Brigotti
M
M
Dipartimento di Patologia sperimentale dell'Università di Bologna, Italy.
Carnicelli
D
D
Alvergna
P
P
Pallanca
A
A
Lorenzetti
R
R
Denaro
M
M
Sperti
S
S
Montanaro
L
L
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D006026:Glycoside Hydrolases / Q000494:pharmacology*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000494:pharmacology*; D010944:Plants / Q000201:enzymology*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D015347:RNA Probes; D012364:RNA, Transfer, Trp / Q000302:isolation & purification / Q000378:metabolism*; D054789:Ribosome Inactivating Proteins, Type 1; D012270:Ribosomes / Q000187:drug effects* / Q000378:metabolism; D016415:Sequence Alignment
D010940:Plant Proteins; D011500:Protein Synthesis Inhibitors; D015347:RNA Probes; D012364:RNA, Transfer, Trp; D054789:Ribosome Inactivating Proteins, Type 1; C025674:GEL protein, Gelonium multiflorum; D006026:Glycoside Hydrolases