A transcription reinitiation intermediate that is stabilized by activator

ABSTRACT High levels of gene transcription by RNA polymerase II depend on high rates of transcription initiation and reinitiation. Initiation requires recruitment of the complete

transcription machinery to a promoter, a process facilitated by activators and chromatin remodelling factors. Reinitiation probably occurs through a different pathway1. After initiation, a

subset of the transcription machinery remains at the promoter, forming a platform for assembly of a second transcription complex2,3,4. Here we describe the isolation of a reinitiation

intermediate that includes transcription factors TFIID, TFIIA, TFIIH, TFIIE and Mediator. This intermediate can act as a scaffold for formation of a functional reinitiation complex.

Formation of this scaffold is dependent on ATP and TFIIH. The scaffold is stabilized in the presence of the activator Gal4–VP16, but not Gal4–AH, suggesting a new role for some activators

and Mediator in promoting high levels of transcription. Access through your institution Buy or subscribe This is a preview of subscription content, access via your institution ACCESS OPTIONS

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RNA POLYMERASE II ASSOCIATES WITH ACTIVE GENES DURING DNA REPLICATION Article 19 July 2023 TRANSCRIPTION FACTORS MODULATE RNA POLYMERASE CONFORMATIONAL EQUILIBRIUM Article Open access 22

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TFIIA is composed of two polypeptide subunits. _J. Biol. Chem._ 266, 19320–19327 (1991). CAS  PubMed  Google Scholar  Download references ACKNOWLEDGEMENTS We thank members of the Hahn and

Reeder laboratories for helpful discussions, and A. Krumm, S. Parkhurst and R. Reeder for comments on the manuscript. We also thank L. Prakash for providing the Rad3ts strain, M. Solomon for

providing the Kin28ts strain, D. Reinberg for TFIIE antibodies and H. Sakurai for providing the Tfa1ts strain and antibodies to Gal11. This work was supported by grants from the NIH to S.H.

and an NIH training grant to N.Y. S.H. is an associate investigator of the Howard Hughes Medical Institute. AUTHOR INFORMATION Author notes * Jeffrey A. Ranish Present address: Institute

for Systems Biology, Seattle, Washington, 98105, USA AUTHORS AND AFFILIATIONS * Division of Basic Sciences, The Fred Hutchinson Cancer Research Center, and Molecular and Cellular Biology

Program, University of Washington, Seattle, 98109, Washington , USA Natalya Yudkovsky, Jeffrey A. Ranish & Steven Hahn * The Howard Hughes Medical Institute , Seattle, 98109, Washington,

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N., Ranish, J. & Hahn, S. A transcription reinitiation intermediate that is stabilized by activator . _Nature_ 408, 225–229 (2000). https://doi.org/10.1038/35041603 Download citation *

Received: 03 August 2000 * Accepted: 25 September 2000 * Issue Date: 09 November 2000 * DOI: https://doi.org/10.1038/35041603 SHARE THIS ARTICLE Anyone you share the following link with will

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