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Science 7 January 1994:
Vol. 263. no. 5143, pp. 81 - 84
DOI: 10.1126/science.8272869
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Articles

Science, Vol 263, Issue 5143, 81-84
Copyright © 1994 by American Association for the Advancement of Science

articles

Mitotic repression of RNA polymerase III transcription in vitro mediated by phosphorylation of a TFIIIB component

JM Gottesfeld, VJ Wolf, T Dang, DJ Forbes, and P Hartl

Department of Molecular Biology, Scripps Research Institute, La Jolla, CA 92037.

Interphase cytosol extracts prepared from Xenopus laevis eggs are active in RNA polymerase III (Pol III) transcription. Addition of recombinant B1 cyclin to these extracts activates mitotic protein kinases that repress transcription. Affinity-purified p34cdc2-cyclin B kinase (mitosis-promoting factor) is sufficient to effect this repression in a simplified Pol III transcription system. This mitotic repression involves the direct phosphorylation of a component of the Pol III transcription initiation factor TFIIIB, which consists of the TATA box-binding protein (TBP) and associated Pol III-specific factors. The transcriptional activity of the TFIIIB-TBP fraction can be modulated in vitro by phosphorylation with mitotic kinases and by dephosphorylation with immobilized alkaline phosphatase.


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