In Escherichia coli, p-aminobenzoate (PABA) is synthesized from chorismate and glutamine in two steps. Aminodeoxychorismate synthase components I and II, encoded by pabB and pabA, respectively, convert chorismate and glutamine to 4-amino-4-deoxychorismate (ADC) and glutamate, respectively. ADC lyase, encoded by pabC, converts ADC to PABA and pyruvate. We reported that pabC had been cloned and mapped to 25 min on the E. coli chromosome (J. M. Green and B. P. Nichols, J. Biol. Chem. 266:12971-12975, 1991). Here we report the nucleotide sequence of pabC, including a portion of a sequence of a downstream open reading frame that may be cotranscribed with pabC. A disruption of pabC was constructed and transferred to the chromosome, and the pabC mutant strain required PABA for growth. The deduced amino acid sequence of ADC lyase is similar to those of Bacillus subtilis PabC and a number of amino acid transaminases. Aminodeoxychorismate lyase purified from a strain harboring an overproducing plasmid was shown to contain pyridoxal phosphate as a cofactor. This finding explains the similarity to the transaminases, which also contain pyridoxal phosphate. Expression studies revealed the size of the pabC gene product to be approximately 30 kDa, in agreement with that predicted by the nucleotide sequence data and approximately half the native molecular mass, suggesting that the native enzyme is dimeric.
A 4589 bp DNA segment containing the Escherichia coli panBCD gene cluster was sequenced, and found to contain 6 complete open reading frames. panB, panC, and panD were identified by subcloning and insertional mutagenesis. The orientation of panD was also confirmed by orientation-specific expression of asparate-1-decarboxylase. panB and panC lie adjacent to one another, but are separated from panD by orf3, which is oriented in the opposite direction. Interruptions in the remaining open reading frames did not affect growth on glucose-minimal medium. No significant similarity to sequences in databases was found for orf1 and orf2. Orf3 contained extensive similarity to reading frames defined by E. coli yjiP, yjiQ, yhgA, and yafD. The function of these amino acid sequences is as yet undefined.
A 4589 bp DNA segment containing the Escherichia coli panBCD gene cluster was sequenced, and found to contain 6 complete open reading frames. panB, panC, and panD were identified by subcloning and insertional mutagenesis. The orientation of panD was also confirmed by orientation‐specific expression of aspartate‐1‐decarboxylase. panB and panC lie adjacent to one another, but are separated from panD by orf3, which is oriented in the opposite direction. Interruptions in the remaining open reading frames did not affect growth on glucose‐minimal medium. No significant similarity to sequences in databases was found for orfl and orf2. Orf3 contained extensive similarity to reading frames defined by E. colib yjiP, yjiQ, yhg A, and yafD. The function of these amino acid sequences is as yet undefined.
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