Nucleotide sequence of the adi gene, which encodes the biodegradative acid-induced arginine decarboxylase of Escherichia coli.
Arginine decarboxylase (encoded by adi) is induced under conditions of acidic pH, anaerobiosis, and rich medium. The DNA sequence of a 3-kb fragment of the Escherichia coli chromosome encoding biodegradative arginine decarboxylase was determined. This sequence encodes a protein of 755 amino acids with a molecular size of 84,420 daltons. The molecular weight and predicted Adi amino acid composition agree with those found in earlier work. The amino acid sequence of arginine decarboxylase showed homology to those of three other decarboxylases of E. coli: (i) CadA, encoding lysine decarboxylase; (ii) SpeC, encoding biosynthetic ornithine decarboxylase; and (iii) SpeF, encoding biodegradative ornithine decarboxylase and the lysine decarboxylase of Hafnia alvei. Unlike SpeC and SpeF, Adi is not similar to the biosynthetic arginine decarboxylase, SpeA. adi is also dissimilar to cadA and speF in that it does not appear to be part of an operon containing a metabolically related transport protein, indicating that it represents a new type of biodegradative decarboxylase regulation. Transcriptional fusions between fragments upstream of adi and lacZ, primer extension, and site-directed mutagenesis experiments defined the pH-regulated promoter. Deletion analysis of the upstream region and cloning of fragments to make adi::lacZ protein fusion implicated a region beyond an upstream SspI site in pH regulation. Induction of adi in the presence of sublethal concentrations of novobiocin or coumermycin A1, inhibitors of DNA gyrase, was dramatically decreased, indicating that DNA supercoiling is involved in adi expression. These results and those of promoter structure studies indicated that acid regulation of adi may involve a mechanism different from that of acid regulation of cad.
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Department of Biochemistry and Cell Biology, Rice University, Houston, Texas 77251.
Bennett
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D000595:Amino Acid Sequence; D049933:Aminocoumarins; D001483:Base Sequence; D001673:Biodegradation, Environmental; D002262:Carboxy-Lyases / Q000235:genetics* / Q000378:metabolism; D002876:Chromosomes, Bacterial; D003374:Coumarins / Q000494:pharmacology; D004250:DNA Topoisomerases, Type II / Q000378:metabolism; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005798:Genes, Bacterial; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009675:Novobiocin / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D017386:Sequence Homology, Amino Acid
D049933:Aminocoumarins; D003374:Coumarins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D009675:Novobiocin; D002262:Carboxy-Lyases; C019722:arginine decarboxylase; D004250:DNA Topoisomerases, Type II; C004628:coumermycin