The binding of streptomycin to ribonucleotides

Incubation of streptomycin (SM) with [32P]5'-ribonucleotides at pH 7.0 produces fractions that migrate towards the cathode in high voltage electrophoresis (HVE) separations at pH 3.5. SM appears to interact with pG, pA and pC but not with pU. The appearance of these [32P]-labeled fractions is dependent on incubation time and SM concentration. Incubation of nucleotides with dihydrostreptomycin (DSM) or SM reduced with sodium cyanoborohydride (NaBH3CN) at pH 5.0, does not produce detectable changes in [32P] nucleotide mobility on HVE; however, incubation with SM reduced with NaBH3CN at pH 7.0 does produce [32P]-labeled fractions migrating with a net positive charge. Elution of [32P]-labeled material migrating towards the cathode from SM-5'-nucleotide incubations and re-electrophoresis results in nucleotides migrating with pG, pA and pC markers. These data indicate a reversible interaction between the SM-streptose aldehyde and amino-group containing nucleotides. This type of interaction may form an additional binding site for SM to RNA, relative to DSM.