SYNTHESIS OF 2,6-DIAZIDO-9-(BETA-D-RIBOFURANOSYL)PURINE 3',5'-BISPHOSPHATE - INCORPORATION INTO TRANSFER-RNA AND PHOTOCHEMICAL LABELING OF ESCHERICHIA-COLI RIBOSOMES

2,6-Diazido-9-(beta-D-ribofuranosyl)purine was prepared by the reactio n of 2,6-dichloro-9-(beta-D-ribofuranosyl)purine with sodium azide. Th e nucleoside was bisphosphorylated with pyrophosphoryl chloride to for m 2,6-diazido-9-(beta-D-ribofuranosyl)purine 3',5'-bisphosphate. This product was labeled with P-32 using T4 polynucleotide kinase to exchan ge the 5' phosphate with the gamma phosphate of ]gamma-P-32]ATP. When yeast tRNA(Phe) containing 2,6-diazido-9-(beta-D-ribofuranosyl)purine at the 3' terminus was bound to the P site of the Escherichia coli rib osome in the presence of poly(U) and irradiated with 300-nm light, the photoreactive tRNA derivative became cross-linked exclusively to the 50S subunit. The label was attached to proteins L27 and L33 as well as to the 23S rRNA.