Mechanism of DmS-II-mediated pause suppression by Drosophila RNA polymerase II.
Transcription elongation factor S-II mediates nascent transcript cleavage by RNA polymerase II (Reines, D. (1992) J. Biol. Chem. 267, 3795-3800). We have examined the mechanism of action of the Drosophila S-II analog, DmS-II, in a defined transcription system. Our results show that DmS-II is necessary and sufficient to activate nascent transcript cleavage by RNA polymerase II during transcription of a dC-tailed template. The pattern of transcripts resulting from prolonged action by DmS-II indicates that there are kinetic barriers to transcript shortening. During the cleavage reaction, the polymerase remains in register with the template strand and generates mainly nucleotide dimers. The ability of DmS-II to mediate transcript shortening resides in the carboxyl-terminal half of the protein. Our results support a model for pause suppression in which DmS-II binds to the paused polymerase, causes one cleavage event and is then released from the complex. Elongation by the polymerase then allows a second encounter with the pause site and a second chance of passing the site. Complete pause suppression may require multiple transcript shortening events for some polymerase molecules.
Guo
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Department of Biochemistry, University of Iowa, Iowa City 52242.
Price
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DH
D000818:Animals; D001665:Binding Sites; D004330:Drosophila / Q000201:enzymology*; D029721:Drosophila Proteins; D007700:Kinetics; D009711:Nucleotides / Q000378:metabolism / Q000494:pharmacology; D012313:RNA / Q000378:metabolism; D012319:RNA Polymerase II / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D016914:Ribonuclease H / Q000378:metabolism; D013698:Templates, Genetic; D014157:Transcription Factors / Q000378:metabolism / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects*
D029721:Drosophila Proteins; D009711:Nucleotides; D011994:Recombinant Proteins; C082903:TfIIS protein, Drosophila; D014157:Transcription Factors; D012313:RNA; D012319:RNA Polymerase II; D016914:Ribonuclease H