Isolation of the putP gene of Corynebacterium glutamicum and characterization of a low-affinity uptake system for compatible solutes

Peter, Heidi; Bader, Andrea; Burkovski, Andreas; Lambert, Camille; Krämer, Reinhard

doi:10.1007/s002030050480

Cited by 28 publications

(31 citation statements)

References 44 publications

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“…Previous tests of this hypothesis have yielded mixed results [14,17,19,20,21,22,27,38,39,41], and the results of our study must be added to those that do not support this hypothesis. This lack of support for the trade-off hypothesis is consistent with recent discoveries in several microbes of both high-and low-affinity uptake systems for a variety of specific nutrients [6,24,31,34,42]. If such dual systems are common in natural populations of bacteria, this would argue against the necessity of a strong trade-off.…”

Section: Discussionsupporting

confidence: 80%

Application of Traditional and Phylogenetically Based Comparative Methods to Test for a Trade-off in Bacterial Growth Rate at Low versus High Substrate Concentration

Velicer¹,

Schmidt²,

Lenski³

1999

Microb Ecol

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A B S T R A C TIt is often hypothesized that those organisms that are superior competitors for sparse resources fare poorly in competition for abundant resources, and vice versa. If there is indeed such a systematic trade-off, then this has important implications for the choice of bacterial strains in bioremediation and other applications. We studied seven bacterial strains that can grow on either 2,4-dichlorophenoxyacetate (2,4-D) or succinate as a sole source of carbon. Growth rates were measured on each substrate at both low (5 µg/ml) and high (500 µg/ml) concentrations. We used two different methods to test the significance of correlations among growth rates, a traditional method that treats each strain as an independent observation and a newer method that takes into account phylogenetic relationships between strains, thereby avoiding spurious correlations caused by a lack of statistical independence of strains. In both 2,4-D and succinate, we observed significant positive correlations between growth rates measured at high and low substrate concentrations by the traditional comparative method. No significant correlations were detected after adjusting for the phylogenetic relationships among the strains. In neither case did we observe the negative correlation expected from a trade-off between growth rates at high and low substrate levels.

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Section: Discussionsupporting

confidence: 80%

Application of Traditional and Phylogenetically Based Comparative Methods to Test for a Trade-off in Bacterial Growth Rate at Low versus High Substrate Concentration

Velicer¹,

Schmidt²,

Lenski³

1999

Microb Ecol

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“…by reducing the expression of cg0834 during the SOS response of C. glutamicum. The putP gene encodes a well-characterized proline/sodium symporter of C. glutamicum that is responsible for proline uptake for anabolic purposes (Peter et al, 1997).…”

Section: Is Lexa Involved In Transcriptional Activation Of Sos Gene Ementioning

confidence: 99%

Genetic makeup of the Corynebacterium glutamicum LexA regulon deduced from comparative transcriptomics and in vitro DNA band shift assays

et al. 2009

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The lexA gene of Corynebacterium glutamicum ATCC 13032 was deleted to create the mutant strain C. glutamicum NJ2114, which has an elongated cell morphology and an increased doubling time. To characterize the SOS regulon in C. glutamicum, the transcriptomes of NJ2114 and a DNA-damage-induced wild-type strain were compared with that of a wild-type control using DNA microarray hybridization. The expression data were combined with bioinformatic pattern searches for LexA binding sites, leading to the detection of 46 potential SOS boxes located upstream of differentially expressed transcription units. Binding of a hexahistidyl-tagged LexA protein to 40 double-stranded oligonucleotides containing the potential SOS boxes was demonstrated in vitro by DNA band shift assays. It turned out that LexA binds not only to SOS boxes in the promoteroperator region of upregulated genes, but also to SOS boxes detected upstream of downregulated genes. These results demonstrated that LexA controls directly the expression of at least 48 SOS genes organized in 36 transcription units. The deduced genes encode a variety of physiological functions, many of them involved in DNA repair and survival after DNA damage, but nearly half of them have hitherto unknown functions. Alignment of the LexA binding sites allowed the corynebacterial SOS box consensus sequence TcGAA(a/c)AnnTGTtCGA to be deduced. Furthermore, the common intergenic region of lexA and the differentially expressed divS-nrdR operon, encoding a cell division suppressor and a regulator of deoxyribonucleotide biosynthesis, was characterized in detail. Promoter mapping revealed differences in divS-nrdR expression during SOS response and normal growth conditions. One of the four LexA binding sites detected in the intergenic region is involved in regulating divS-nrdR transcription, whereas the other sites are apparently used for negative autoregulation of lexA expression.

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“…For gram-negative bacterial species, the regulation of the putP gene has been examined (18,30,32,38), but only limited analyses of putP regulation have been done for gram-positive bacterial species (36,48,50). Presently, no study has looked at the transcriptional regulation of the putP gene in S. aureus.…”

mentioning

confidence: 99%

Transcriptional Activation of the Staphylococcus aureus putP Gene by Low-Proline-High Osmotic Conditions and during Infection of Murine and Human Tissues

2006

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Staphylococcus aureus can grow virtually anywhere in the human body but needs to import proline through low-and high-affinity proline transporters to survive. This study examined the regulation of the S. aureus putP gene, which encodes a high-affinity proline permease. putP::lacZ and putP::lux transcriptional fusions were constructed and integrated into the genomes of several S. aureus strains. Enzyme activity was measured after growth in media with various osmolyte concentrations. As osmolarity rose, putP expression increased, with a plateau at 2 M for NaCl in strain LL3-1. Proline concentrations as low as 17.4 M activated expression of the putP gene. The putP::lux fusion was also integrated into the genomes of S. aureus strains that were either SigB inactive (LL3-1, 8325-4, and SH1003) or SigB active (Newman and SH1000). SigB inactive strains showed increased putP gene expression as NaCl concentrations rose, whereas SigB active strains displayed a dramatic decrease in putP expression, suggesting that the alternative sigma factor B plays a negative role in putP regulation. Mice inoculated with S. aureus strains containing the putP::lux fusion exhibited up to a 715-fold increase in putP expression, although levels in the various murine organs differed. Moreover, urine from human patients infected with S. aureus showed elevated putP levels by use of a PCR procedure, whereas blood and some abscess material had no significant increase. Thus, putP is transcriptionally activated by a lowproline and high osmotic environment both in growth media and in murine or human clinical specimens.

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Isolation of the putP gene of Corynebacterium glutamicum and characterization of a low-affinity uptake system for compatible solutes

Cited by 28 publications

References 44 publications

Application of Traditional and Phylogenetically Based Comparative Methods to Test for a Trade-off in Bacterial Growth Rate at Low versus High Substrate Concentration

Application of Traditional and Phylogenetically Based Comparative Methods to Test for a Trade-off in Bacterial Growth Rate at Low versus High Substrate Concentration

Genetic makeup of the Corynebacterium glutamicum LexA regulon deduced from comparative transcriptomics and in vitro DNA band shift assays

Transcriptional Activation of the Staphylococcus aureus putP Gene by Low-Proline-High Osmotic Conditions and during Infection of Murine and Human Tissues

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