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(Received for publication, June 19, 1996, and in revised form, August 13, 1996)
From the Molecular Virology Section, Laboratory of Molecular
Microbiology, NIAID, National Institutes of Health,
Bethesda, Maryland 20892-0460
The carboxyl-terminal domain (CTD) of RNA
polymerase (RNAP) II contains multiple repeats with a heptapeptide
consensus: Tyr-Ser-Pro-Thr-Ser-Pro-Ser. It has been proposed that
phosphorylation of this CTD facilitates clearance and elongation of
transcription complexes initiated at the promoters. However, not all
transcribed promoters require RNAP II with full-length CTD.
Furthermore, different activators can promote capably the
transcriptional activity of polymerase II mutants deleted in the CTD.
Thus, the role of the RNAP II CTD in transcription and in response to
activators remains incompletely understood. To study the role of CTD in
the regulated transcription of human retroviruses human-T cell
lymphotropic virus I and human immunodeficiency virus 1, we used an
Volume 271, Number 44,
Issue of November 1, 1996
pp. 27888-27894
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
-amanitin-resistant system developed previously (Gerber, H. P.,
Hagmann, M., Seipel, K., Georgiev, O., West, M. A., Litingtung, Y.,
Schaffner, W., and Corden, J. L. (1995) Nature 374, 660-662). We found that transcription directed by the human T-cell
lymphotropic virus I activator protein Tax was strongly promoted by
CTD-deficient RNA polymerase II. By contrast, the human
immunodeficiency virus 1 activator Tat, which is recruited to the
promoter by tethering to a nascent leader RNA, requires CTD-containing
polymerase II for transcriptional activity. Biochemically, we
characterized that Tat associated with a cellular CTD kinase activity,
whereas Tax did not. Concordantly, we found that cellular transcription
factor Sp1, which can activate CTD-deficient polymerase II with an
efficiency similar to Tax, also failed to bind a CTD kinase. Taken
together, these observations address mechanistic corollaries between
activators with(out) a linked CTD kinase and regulated transcription by
RNA polymerase II moieties with(out) a CTD.
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