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KMID : 0380219960290010092
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Journal of Biochemistry and Molecular Biology
1996 Volume.29 No. 1 p.92 ~ p.97
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Site-Directed Mutation Effect of the Symmetry Region at the mRNA 5'-end of Escherichia coli aeg-46.5 Gene
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Ahn Ju-Hyuk
Choe Mu-Hyeon
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Abstract
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The age-46.5 gene of Escherichia coli is induced by nitrate ion and regulated by Fnr, NarL, and NarP during anaerobic growth. aeg-46.5::lacZ fusion gene shows its maximum expression in narL host after two hours of aerobic to anaerobic switch in M9-Glc-nitrate medium. Fnr and NarP act as positive regulators, and NarL acts as a negative regulator. The control region of the aeg-46.5 was identified and the binding sites of regulator proteins have been predicted (Reznikoff and Choe (1993)). It has two symmetry regions. One is located at -52~-37 bp from the anaerobic mRNA 5'-end, which is the binding site of NarL and NarP. The other is located at +37~+56 bp from the 5'-end of mRNA. In this study, the downstream symmetry region from the mRNA 5'-end was investigated by site-directed mutagenesis. The destruction of the symmetry region increases the expression level of aeg-46.5. We propose that the symmetry region interferes with the expression of aeg-46.5 possibly by forming a stem-and-loop structure.
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KEYWORD
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aeg-46.5, anaerobically expressed gene, NarL, site-directed mutagenesis, symmetry region
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