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Nucleic Acids Research, 1995, Vol. 23, No. 9 1551-1556
© 1995

MOLECULAR BIOLOGY

Interaction of Xenopus TFIIIC with a 5S RNA gene

Michael R. Sturges+, Magdalena Bartilson§ and Lawrence J. Peck*

Sinsheimer Laboratories, University of California at Santa Cruz Santa Cruz, CA 95064, USA

* To whom correspondence should be addressed

Received December 19, 1994. Revised March 17, 1995. Accepted March 17, 1995.

Using fractionated Xenopus transcription factors we have identified and characterized a unique protein-DNA complex formed between TFIIIA, TFIIIC and a 5S RNA gene. The formation of this complex was blocked by specific competitor DNAs and by the inactivatlon of TFIIIC using two different methods. In addition, TFIIIC activity was retained when the complexes were affinity purified using a reverslbly Immobilized DNA template. The TFIII(A+C)-5S RNA gene complex has a distlnct electrophoretic mobility on band-shift gels and a unique DNase I footprint. The characteristic feature of the DNase I footprint is a TFIIIC-dependent extension of the TFIIIA footprint an additional 25 bp toward the 5' end of the gene. This Indicates a direct interaction between Xenopus TFIIIC and the template DNA.

+Department of Pediatrics, Oregon Health Sciences University, Portland, OR 97201, USA

§Protein Design Labs, Mountain View, CA 94043, USA


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