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A more recent version of this article appeared on August 4, 2000
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M001883200v1
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Papers In Press, published online ahead of print May 23, 2000
J. Biol. Chem, 10.1074/jbc.M001883200
Submitted on March 4, 2000
Revised on May 22, 2000
Accepted on May 23, 2000

Conditional expression of RNA polymerase II in mammalian cells: deletion of the carboxy-terminal domain of the large subunit affects early steps in transcription

Mark Meininghaus, Rob D. Chapman, Manuela Horndasch, and Dirk Eick

Institute of Clinical Molecular Biology and Tumor Genetics, GSF, Munich D-81377

Corresponding Author: eick{at}gsf.de

The carboxy-terminal domain (CTD) of the large subunit of mammalian RNA polymerase II contains 52 repeats of a heptapeptide which is the target of a variety of kinases. The hyperphosphorylated CTD recruits important factors for mRNA capping, splicing and 3'-processing. The role of the CTD for the transcription process in vivo, however, is not yet clear. We have conditionally expressed an a-amanitin-resistant large subunit with an almost entirely deleted CTD (LS*D5) in B-cells. These cells have a defect in global transcription of cellular genes in the presence of a-amanitin. Moreover, pol II harboring LS*D5 failed to transcribe up to the promoter-proximal pause sites in the hsp70A and c-fos gene promoters. The results indicate that the CTD is already required for steps that occur before promoter-proximal pausing and maturation of mRNA.


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