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Transcriptional termination and coupled polyadenylation in vitro
Author(s) -
Yonaha Masatomo,
Proudfoot Nick J.
Publication year - 2000
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/19.14.3770
Subject(s) - polyadenylation , biology , cleavage and polyadenylation specificity factor , transcription (linguistics) , rna polymerase ii , cleavage factor , microbiology and biotechnology , post transcriptional modification , terminator (solar) , in vitro , genetics , rna , rna splicing , gene expression , promoter , gene , physics , ionosphere , linguistics , philosophy , astronomy
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.