Molecular dissection of mammalian RNA polymerase II transcriptional termination.
Transcriptional termination of mammalian RNA polymerase II (Pol II) is an essential but little-understood step in protein-coding gene expression. Mechanistically, termination by all DNA-dependent RNA polymerases can be reduced to two steps, namely release of the RNA transcript and release of the DNA template. Using a simple nuclear fractionation procedure, we have monitored transcript and template release in the context of both natural and artificial Pol II terminator sequences. We describe the timing and relationship between these events and in so doing establish the roles of the poly(A) signal, cotranscriptional RNA cleavage events, and 5'-3' exonucleolytic RNA degradation in the mammalian Pol II termination process.
West
Steven
S
Sir William Dunn School of Pathology, South Parks Road, Oxford OX1 3RE, UK.
Proudfoot
Nicholas J
NJ
Dye
Michael J
MJ
D020413:3' Untranslated Regions; D000818:Animals; D005786:Gene Expression Regulation; D005914:Globins / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D012313:RNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism*; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D014158:Transcription, Genetic
D020413:3' Untranslated Regions; D012322:RNA Precursors; D012333:RNA, Messenger; D012313:RNA; D005914:Globins; D012319:RNA Polymerase II