Dichlorobenzimidazole-riboside inhibition of nuclear RNA accumulation initiated with gamma-thio analogues of ATP and GTP

Abstract

The gamma-thio analogues of ATP and GTP, ATP[S] and GTP[S], have been used as affinity probes to measure RNA synthesis initiated in vitro in L cell nuclei. 5,6-Dichlororibofuranosylbenzimidazole was found to inhibit total RNA synthesis in vitro and the amount bound by mercury-affinity chromatography. Initiation in vitro was also shown by [35S]ATP[S] and [35S]GTP[S] incorporation in RNA molecules with a larger size distribution. Although 66% of RNA molecules labeled with [35S]GTP[S] and 74% of those labeled with [35S]ATP[S] were 3-12 S n size, the remainder of the label was recovered in RNA molecules larger than 12 S and a significant portion in RNA larger tha 18 S. The specificity of the initiation process seems to be indicated by the finding of molecules of the size of pre-tRNA on gel electrophoresis which were labeled with [35S]GTP[S] but not [35S]ATP[S] under our experimental conditions. 5,6-Dichlororibofuranosylbenzimidazole severely inhibited the incorporation of [35S]GTP[S] and [35S]AT[[S] by all size classes, indicating that it can decrease accumulation of RNA initiated in vitro in the L cell nuclear system.