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Functional analysis of Drosophila factor 5 (TFIIF), a general transcription factor
Journal article   Open access   Peer reviewed

Functional analysis of Drosophila factor 5 (TFIIF), a general transcription factor

Daniel D Kephart, Bo Qing Wang, Zachary F Burton and David H Price
The Journal of biological chemistry, Vol.269(18), pp.13536-13543
1994
DOI: 10.1016/S0021-9258(17)36864-3
PMID: 8175788
url
https://doi.org/10.1016/S0021-9258(17)36864-3View
Published (Version of record) Open Access

Abstract

Factor 5 is a Drosophila RNA polymerase II initiation factor that also affects the elongation phase of transcription. We have used a cDNA encoding the large subunit of factor 5 (F5a) to produce recombinant F5a (rF5a). Antibodies directed against peptides deduced from the sequence of the F5a cDNA recognized rF5a and the large subunit of factor 5 purified from Kc cells. A chimeric human/fly factor composed of the small subunit of human TFIIF (RAP30) and rF5a stimulated elongation by Drosophila RNA polymerase II when assayed using a dC-tailed template. In addition, the chimeric human/fly factor functioned during initiation in either the Drosophila or human system. Therefore, the structure of the large subunit of TFIIF is sufficiently conserved from human to fly to allow functional interaction with both the small subunit of TFIIF and RNA polymerase II from either species. Analysis of deletion mutants of F5a indicated that almost all of the protein was required for initiation while only the NH2-terminal region was required for stimulating transcriptional elongation. A comparison of our results with those obtained with RAP74 suggest that the carboxyl terminal region of the protein may be involved in interactions with RNA polymerase II or other factors during initiation.
Molecular Genetics Fundamental and applied biological sciences. Psychology Biological and medical sciences Transcription. Transcription factor. Splicing. Rna processing Molecular and cellular biology

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