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Nucleic Acids Research, 1994, Vol. 22, No. 23 4958-4962
© 1994

MOLECULAR BIOLOGY

RNA polymerase II phosphorylation: uncoupling from GAL4-VP16 directed open complex formation and transcription in a reconstituted system

Ying Jiang and Jay D. Gralla*

Department of Chemistry and Biochemistry and the Molecular Biology Institute, University of California Los Angeles, 405 Hilgard Avenue, Los Angeles, CA 90024-1569

*To whom correspondence should be addressed

Received August 17, 1994. Revised October 17, 1994. Accepted October 17, 1994.

An activated transcription system was constructed using substantially purified liver factors, Hela TFIID and GAL4-VP16. The system was used to study the relationship between RNA polymerase II large subunit phosphorylation and other ATP-dependent processes occurring during activated transcription. When Cterminal domain (CTD) kinase activity was inhibited, activator dependent open promoter complex formation proceeded normally. These open complexes could function to produce RNA in the absence of CTD phosphorylation, although the level of RNA produced was changed somewhat. The results demonstrate that RNA polymerase II CTD phosphorylation is not generally required for the formation of activatordependent, functional open promoter complexes. Taken together with prior results the experiments suggest that a requirement for CTD phosphorylation may be situation-dependent and thus serve a regulatory function.


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