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Molecular and Cellular Biology, October 2000, p. 7438-7449, Vol. 20, No. 20
Banting and Best Department of Medical
Research1 and Department of Molecular
and Medical Genetics,2 University of
Toronto, Toronto, Ontario M5G 1L6, and Department of Biological
Sciences, Boehringer Ingelheim (Canada) Limited, Laval, Quebec H7S
2G5,4 Canada, and Department of
Biochemistry and Molecular Biology, University of Georgia, Athens,
Georgia 306023
Received 13 June 2000/Returned for modification 16 July
2000/Accepted 31 July 2000
Transcription by RNA polymerase II is accompanied by cyclic
phosphorylation and dephosphorylation of the carboxy-terminal heptapeptide repeat domain (CTD) of its largest subunit. We have used
deletion and point mutations in Fcp1p, a TFIIF-interacting CTD
phosphatase, to show that the integrity of its BRCT domain, like
that of its catalytic domain, is important for cell viability, mRNA synthesis, and CTD dephosphorylation in vivo.
Although regions of Fcp1p carboxy terminal to its BRCT domain and at
its amino terminus were not essential for viability, deletion of either of these regions affected the phosphorylation state of the CTD. Two portions of this carboxy-terminal region of Fcp1p bound
directly to the first cyclin-like repeat in the core domain of the
general transcription factor TFIIB, as well as to the RAP74 subunit of TFIIF. These regulatory interactions with Fcp1p involved closely related amino acid sequence motifs in TFIIB and RAP74. Mutating the
Fcp1p-binding motif KEFGK in the RAP74 (Tfg1p) subunit of TFIIF to
EEFGE led to both synthetic phenotypes in certain fcp1 tfg1 double mutants and a reduced ability of Fcp1p to activate transcription when it is artificially tethered to a promoter. These results suggest strongly that this KEFGK motif in RAP74 mediates
its interaction with Fcp1p in vivo.
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
A Motif Shared by TFIIF and TFIIB Mediates Their Interaction
with the RNA Polymerase II Carboxy-Terminal Domain Phosphatase
Fcp1p in Saccharomyces cerevisiae
*
Corresponding author. Mailing address: Banting and Best
Department of Medical Research, University of Toronto, 112 College St.,
Rm. 210, Toronto, Ontario, Canada M5G 1L6. Phone: (416) 978-4141. Fax:
(416) 978-8528. E-mail: jack.greenblatt{at}utoronto.ca.
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