1887

Abstract

Proline utilization (Put) systems have been described in a number of bacteria; however, the importance and functionality of the Put system in the intracellular pathogen has not been explored. Generally, bacterial Put systems are composed of the bifunctional enzyme proline dehydrogenase PutA and its transcriptional activator PutR. Here, we demonstrate that the genes () and () are critical for the chronic infection of mice by , but and are not required for the survival and replication of the bacteria in naive macrophages. Additionally, experiments revealed that is necessary for the ability of the bacteria to withstand oxidative stress, as a Δ deletion strain is hypersensitive to hydrogen peroxide exposure. Quantitative reverse transcription-PCR and transcriptional reporter studies revealed that PutR acts as a transcriptional activator of in , and electrophoretic mobility shift assays confirmed that PutR binds directly to the promoter region. Biochemical analyses demonstrated that a purified recombinant PutA protein possesses quintessential proline dehydrogenase activity, as PutA is capable of catalysing the conversion of proline to glutamate. Altogether, these data are the first to reveal that the Put system plays a significant role in the ability of to replicate and survive within its host, as well as to describe the genetic regulation and biochemical activity of the Put system in .

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2017-07-01
2024-03-28
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