An RNA polymerase II complex containing capping enzymes and viral oncoproteins

The aim of this work was to identify proteins from Adenovirus 2-infected HeLa cell extracts that interact with the carboxyl-terminal domain of the largest subunit of RNA polymerase II. First, a mammalian RNA polymerase II complex was isolated from Adenovirus 2-infected HeLa cell extracts by affinity chromatography against the carboxyl-terminal domain of the largest subunit of RNA polymerase II, followed by chromatography on a Mono S fast protein liquid chromatographic column. Second, the isolated complex was further characterized by Western blot analysis, the formation of a GMP-protein complex, and transcriptional activity. The isolated complex contains general transcription factors, chromatin-remodeling factors, histone acetyltransferases, Srbs, capping enzymes, and E1A viral oncoproteins. The RNA polymerase II complex is active in transcription when supplemented with recombinant transcription factor IIB.