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Science. 1993 Oct 8;262(5131):244-8.

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

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1
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.

Abstract

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.

PMID:
8211143
[Indexed for MEDLINE]
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