Transcriptional termination and coupled polyadenylation in vitro.
Using a coupled, in vitro transcription and polyadenylation system we have investigated the molecular mechanism of transcriptional termination by RNA polymerase II (PolII). We showed previously that specific G-rich sequences pause transcription and then activate polyadenylation. We show that physiological pause sites activate polyadenylation in our in vitro system. We also investigate the mechanism of PolII transcriptional termination, and show that these transcripts are either directly released from the transcription complex or are 3' end processed while still attached to the complex. We also show that 3' product (generated by cleavage/polyadenylation) remains associated with the transcription complex, but is rapidly degraded on it.
Yonaha
M
M
Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK.
Proudfoot
N J
NJ
D003175:Complement C2 / Q000235:genetics; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins; D006801:Humans; D009838:Oligodeoxyribonucleotides / Q000235:genetics / Q000378:metabolism; D011061:Poly A / Q000235:genetics / Q000378:metabolism; D012319:RNA Polymerase II / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D020218:Response Elements / Q000235:genetics; D012521:Sarcosine / Q000031:analogs & derivatives / Q000494:pharmacology; D013698:Templates, Genetic; D013728:Terminator Regions, Genetic / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects / Q000235:genetics*
D003175:Complement C2; D004268:DNA-Binding Proteins; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D014157:Transcription Factors; C076035:c-MYC-associated zinc finger protein; D011061:Poly A; C025231:sarkosyl; D004247:DNA; D012319:RNA Polymerase II; D012521:Sarcosine