1887

Abstract

experiments have shown that the genes of the pyrimidine biosynthetic pathway of , the genes, are regulated by a transcriptional attenuation mechanism. Specific regulatory sequences (binding loops, BLs) are located within three untranslated leader sequences at the beginning of mRNA. These binding loops, BL1, BL2 and BL3, act as anti-antiterminators of transcription when stabilized by the regulator protein PyrR. In this work, the interaction of PyrR with BL1, BL2 and BL3 was qualitatively and quantitatively analysed using the yeast three-hybrid system. The results indicate that PyrR specifically binds to BL1, BL2 and BL3. Furthermore, the data suggest that the strength of interaction between PyrR and the three different BLs is within the same dimension. The yeast three-hybrid system also proved to be useful for the rapid analysis of structural requirements for PyrR–BL binding. Point mutations within the predicted critical regions of BL1, BL2 and BL3 led to drastically reduced binding of PyrR. In summary, it is shown that the yeast three-hybrid system is well suited to qualitatively and quantitatively analyse bacterial regulatory systems that are based on factor-independent transcriptional attenuation.

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2007-03-01
2019-10-22
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