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Science 8 October 1993:
Vol. 262. no. 5131, pp. 244 - 248
DOI: 10.1126/science.8211143
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Articles

Science, Vol 262, Issue 5131, 244-248
Copyright © 1993 by American Association for the Advancement of Science

articles

Regional codon randomization: defining a TATA-binding protein surface required for RNA polymerase III transcription

BP Cormack and K Struhl

Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.

The TATA-binding protein (TBP) is required for transcription by all three nuclear RNA polymerases. TBP was subjected to regional codon randomization, a codon-based mutagenesis method that generates complex yet compact protein libraries. Analysis of 186 temperature-sensitive TBP mutants yielded 65 specifically defective in transcription by RNA polymerase III (Pol III). These mutants map to a limited TBP surface that may interact with Tds4, a component of the Pol III transcription factor TFIIIB. Strains that contain the Pol III-defective derivatives have increased amounts of messenger RNA, which suggests that competition among TBP-interacting factors for limiting quantities of TBP determines the ratio of Pol II and Pol III transcription in vivo.


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