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Nucleic Acids Research, 2000, Vol. 28, No. 20 3935-3942
© 2000 Oxford University Press

Transcription efficiency of human polymerase III genes in vitro does not depend on the RNP-forming autoantigen La

Stephan Weser, Michael Bachmann1, Klaus H. Seifart* and Wolfgang Meißner

Institut für Molekularbiologie und Tumorforschung (IMT), Philipps Universität Marburg, Lahnstraße 3, D-35037 Marburg, Germany and 1Oklahoma Medical Research Foundation (OMRF), 323 NE 13th Street, 73105 Oklahoma City, OK, USA

Transcription of class III genes is conducted by multi-protein complexes consisting of polymerase III itself and several transcription factors. We established a reconstituted in vitro transcription system from which the autoantigen La was removed by immunodepletion. This system showed no RNP formation, but was still fully active in transcription. Supplementing such La-free transcription reactions with recombinant La restored the formation of La complexes with the newly synthesised RNA, but did not lead to enhanced transcription efficiency. Furthermore, we developed a technique for the generation and isolation of transcription complexes, assembled from purified transcription factors and isolated by glycerol centrifugation. These complexes were fully competent to re-initiate RNA synthesis but they were not associated with La and their transcription rate could not be stimulated by addition of recombinant La. Therefore, we conclude that La does not act as a human polymerase III transcription factor.

* To whom correspondence should be addressed. Tel: +49 6421 2865013; Fax: +49 6421 2865010; Email: seifart@imt.uni-marburg.de


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