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PMID:23995643

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Citation

'Lyell, NL, Colton, DM, Bose, JL, Tumen-Velasquez, MP, Kimbrough, JH and Stabb, EV (2013) CRP regulates pheromone-mediated bioluminescence at multiple levels in Vibrio fischeri ES114. J. Bacteriol. '

Abstract

Bioluminescence in Vibrio fischeri ES114 is activated by autoinducer pheromones, and this regulation serves as a model for bacterial cell-cell signaling. As in other bacteria, pheromone concentration increases with cell density; however, pheromone synthesis and perception are also modulated in response to environmental stimuli. Previous studies suggested that expression of the pheromone-dependent bioluminescence activator LuxR is regulated in response to glucose by cAMP-receptor protein (CRP). Consistent with this model, we found that bioluminescence in V. fischeri ES114 is modulated by glucose and stimulated by cAMP. In addition, a Δcrp mutant was ∼100-fold dimmer than ES114 and did not increase luminescence in response to added cAMP, even though cells lacking crp were still metabolically capable of producing luminescence. We further discovered that CRP regulates not only luxR, but also the alternative pheromone synthase gene ainS. We found that His-tagged V. fischeri CRP could bind sequences upstream of both luxR and ainS, supporting bioinformatic predictions of direct regulation at both promoters. Luminescence increased in response to cAMP if either the ainS or luxR systems were under native regulation, suggesting cAMP-CRP significantly increases luminescence through both systems. Finally, using transcriptional reporters in transgenic Escherichia coli we elucidated two additional regulatory connections. First, LuxR-independent basal transcription of the luxI promoter was enhanced by CRP. Second, the effect of CRP on the ainS promoter depended on whether the V. fischeri regulatory gene litR was also introduced. These results suggest an integral role for CRP in pheromone signaling that goes beyond sensing cell density.

Links

PubMed Online version:10.1128/JB.00751-13

Keywords


Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

ALIF1:Q5E2H1

enables

GO:0000976: transcription cis-regulatory region binding

ECO:0005632: fluorescence anisotropy evidence used in manual assertion

F

Seeded From UniProt

complete

ALIF1:Q5E2H1

enables

GO:0001216: DNA-binding transcription activator activity

ECO:0005632: fluorescence anisotropy evidence used in manual assertion

F

Seeded From UniProt

complete

ALIF1:Q5E2H1

part_of

GO:0032993: protein-DNA complex

ECO:0005632: fluorescence anisotropy evidence used in manual assertion

C

Seeded From UniProt

complete

ALIF1:Q5E2H1

involved_in

GO:0045893: positive regulation of transcription, DNA-templated

ECO:0005636: green fluorescent protein reporter gene assay evidence used in manual assertion

P

Seeded From UniProt

complete

ALIF1:Q5E2H1

involved_in

GO:0045893: positive regulation of transcription, DNA-templated

ECO:0005660: quantitative polymerase chain reaction evidence used in manual assertion

P

Seeded From UniProt

complete

VIBF1:Q5E2H1

enables

GO:0000976: transcription regulatory region sequence-specific DNA binding

ECO:0005632: fluorescence anisotropy evidence used in manual assertion

F

Seeded From UniProt

complete

VIBF1:Q5E2H1

enables

GO:0001216: DNA-binding transcription activator activity

ECO:0005632: fluorescence anisotropy evidence used in manual assertion

F

Seeded From UniProt

complete

VIBF1:Q5E2H1

part_of

GO:0032993: protein-DNA complex

ECO:0005632: fluorescence anisotropy evidence used in manual assertion

C

Seeded From UniProt

complete

VIBF1:Q5E2H1

involved_in

GO:0045893: positive regulation of transcription, DNA-templated

ECO:0005636: green fluorescent protein reporter gene assay evidence used in manual assertion

P

Seeded From UniProt

complete

VIBF1:Q5E2H1

involved_in

GO:0045893: positive regulation of transcription, DNA-templated

ECO:0005660: quantitative polymerase chain reaction evidence used in manual assertion

P

Seeded From UniProt

complete

VIBF1:Q5E833

involved_in

GO:0045893: positive regulation of transcription, DNA-templated

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

VIBF1:Q5E833

involved_in

GO:0009371: positive regulation of transcription by pheromones

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

VIBF1:Q5E833

GO:0009371 : positive regulation of transcription by pheromones

ECO:0000315:

P

Figure 6. Regulation of both ainS and luxR contributes to the effect of cAMP on luminescence.

complete
CACAO 8765

VIBF1:Q5E833

GO:0008218: bioluminescence

ECO:0000315:

P

Figure 2. Effect of cAMP on luminescence.

complete
CACAO 8766

VIBF1:Q5E833

GO:0043193: positive regulation of gene-specific transcription

ECO:0000315:

P

Figure 4. The luxR and ainS transcripts was decreased by ~250-fold and ~7-fold, respectively in the delta crp mutant.

complete
CACAO 8767

VIBF1:Q5E833

GO:0032583: regulation of gene-specific transcription

ECO:0000315:

P

Figure 2.

complete
CACAO 8768

VIBF1:Q5E833

involved_in

GO:0008218: bioluminescence

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

See also

References

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