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Nucleic Acids Research, 1993, Vol. 21, No. 4 1013-1018
© 1993

MOLECULAR BIOLOGY

Transcription factor IIA stimulates the expression of classical pollll-genes

Wolfgang Meißner, Rudi Holland, Rainer Waldschmidt and Klaus H. Seifart*

Institut für Molekutarbiologie und Tumorforschung der Philipps Universftät Marburg Lahnstr.3, D-3550 Marburg, Germany

* To whom correspondence should be addressed

Received September 25, 1992. Revised January 19, 1993. Accepted January 19, 1993.

Protein fractions containing TFIIA, a transcription factor known to be involved in transcription initiation by RNA polymerase II and 5'-regulated polymerase III genes (e.g. U6), were tested for their role in in vitro transcription of classical pol III genes. These fractions were shown to stimulate a basal transcription system, reconstituted from highly purified fractions hTFIIIB and hTFIIIC. We demonstrate that this stimulating activity isolated from HeLa cells coelutes over at least six chromatographic steps with hTFIIA. Moreover the native molecular mass and the stability of this activity against heat treatment are comparable to those of hTFIIA. Finally we show that recombinant TFIIA from Saccharomyces cerevisiae can substitute for the human factor in pol III transcription in vitro which proves that TFIIA is also involved in the efficient expression of classical pol III genes.


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