Reconstitution of Bacillus subtilis trp attenuation in vitro with TRAP, the trp RNA-binding attenuation protein

P. Babitzke, C. Yanofsky

Research output: Contribution to journalArticlepeer-review

110 Scopus citations

Abstract

We have reconstituted Bacillus subtilis trp attenuation in vitro. Purification of the mtrB gene product (TRAP) to near homogeneity allowed us to demonstrate that addition of this protein plus L-tryptophan to template, RNA polymerase, and nucleoside triphosphates caused transcription termination in the trpEDCFBA leader region. TRAP acts by binding to the nascent transcript and preventing formation of an RNA antiterminator structure, thereby allowing terminator formation and transcription termination. Oligonucleotides complementary to segments of the antiterminator were used to demonstrate that formation of this RNA hairpin was responsible for transcription read-through. TRAP was found to be a 60-kDa multimeric protein composed of identical 6- to 8-kDa subunits, and its elution profile on a chromatographic column did not change in the presence of tryptophan.

Original languageEnglish (US)
Pages (from-to)133-137
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number1
DOIs
StatePublished - 1993

All Science Journal Classification (ASJC) codes

  • General

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