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Volume 271, Number 20, Issue of May 17, 1996 pp. 11703-11709
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
RNA Polymerase II-associated Protein (RAP) 74 Binds Transcription Factor (TF) IIB and Blocks TFIIB-RAP30 Binding

(Received for publication, October 19, 1995; and in revised form, February 6, 1996)

Shi Min Fang,  Zachary F. Burton

A set of deletion mutants of human RNA polymerase II-associated protein (RAP) 30, the small subunit of transcription factor IIF (TFIIF; RAP30/74), was constructed to map functional domains. Mutants were tested for accurate transcriptional activity, RAP74 binding, and TFIIB binding. Transcription assays indicate the importance of both N- and C-terminal sequences for RAP30 function. RAP74 binds to the N-terminal region of RAP30 between amino acids 1 and 98. TFIIB binds to an overlapping region of RAP30, localized to amino acids 1-176 (amino acids 27-152 comprise a minimal binding region). The C-terminal region of RAP74 (amino acids 358-517) binds directly and independently to TFIIB. Interestingly, RAP74 blocks TFIIB-RAP30 binding, both by binding TFIIB and by binding RAP30. When the TFIIF complex is intact, therefore, TFIIB-TFIIF contact is maintained through RAP74. If the TFIIB-RAP30 interaction is physiologically important, the TFIIF complex must dissociate within some transcription complexes.



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