1887

Abstract

PAO1 utilizes 3-guanidinopropionate (3-GP) and 4-guanidinobutyrate (4-GB), which differ in one methylene group only, via distinct enzymes: guanidinopropionase (EC 3.5.3.17; the product) and guanidinobutyrase (EC 3.5.3.7; the product). The authors cloned and characterized the contiguous genes (in that order) responsible for 3-GP utilization, and compared the deduced sequences of their putative protein products, and the potential regulatory mechanisms of , with those of the corresponding genes encoding the 4-GB catabolic system. GpuA and GpuR have similarity to GbuA (49 % identity) and GbuR (a transcription activator of ; 37 % identity), respectively. GpuP resembles PA1418 (58 % identity), which is a putative membrane protein encoded by a potential gene downstream of . These features of the GpuR and GpuP sequences, and the impaired growth of and knockout mutants on 3-GP, support the notion that GpuR and GpuP direct the 3-GP-inducible expression of , and the uptake of 3-GP, respectively. Northern blots of mRNA from 3-GP-induced PAO1 cells revealed three transcripts of , , and and together, suggesting that and each have a 3-GP-responsible promoter, and that some transcription from the promoter is terminated after , or proceeds into . Knockout of abolished 3-GP-dependent synthesis of the transcripts, confirming that GpuR activates transcription from these promoters, with 3-GP as a specific co-inducer. The sequence conservation between the three functional pairs of the Gpu and Gbu proteins, and the absence of in closely related species, imply that the triad genes have co-ordinately evolved from origins common to the counterparts, to establish an independent catabolic system of 3-GP in .

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2005-12-01
2019-11-22
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