Spontaneous mutations affecting transcriptional regulation by protocatechuate in Acinetobacter
D A D'Argenio1, A Segura, P V Bünz
1Department of Molecular, Cellular and Developmental Biology, Yale University, P.O. Box 208103, New Haven, CT 06520-8103, USA.
Abstract:
Positive selection yields Acinetobacter strains with a spontaneous mutation blocking catabolism of protocatechuate. For this study, the growth temperature during selection was lowered to 22 degrees C: growth at 37 degrees C was found to mask the role of the protocatechuate-responsive transcriptional regulator PcaU. The resulting mutants included those with amino acid substitutions useful for understanding PcaU structure and function, a 20-bp deletion whose repeated isolation suggested genetic instability of DNA in the putative PcaU operator, and a large deletion whose phenotype revealed that the supraoperonic cluster of genes for the protocatechuate branch of the beta-ketoadipate pathway extends to genes for the utilization of C(6)-C(10) straight-chain dicarboxylic acids including adipate.
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