Part:BBa_K4140001

TyrR

Part Description

The TyrR protein in Escherichia coli has the ability to both activate and repress the transcription operons necessary for the production and intake of aromatic amino acids (tyrosine, phenylalanine, and tryptophan). As an illustration, the TyrR dimer activates the promoter of the tyrP gene, which codes for a transporter specific to tyrosine, when tyrosine is present

Usage

We took advantage of The TyrR protein properties and it’s ability to control the biosynthesis and intake of aromatic amino acids (tyrosine, phenylalanine, and tryptophan). So we employ it to control the activity of the paroF promoter and TyrP promoter as TyrR dimer activates the TyrP promoter and paroF promoter in the presence of phenylalanine , when tyrosine is more abundant the induction effect on paroF is lost

Literature Characterization

ATPase activity of tyrR:
In this study, TyrR has poor ATPase activity that ranges between 12 and 400 mmol of ATP mol−1 monomer min−1. TyrR-(188–467) posess a specific ATPase activity of 105 mmol of ATP mol−1 monomer min−1, about five times the value for TyrR, which was found in this study to be 20 mmol of ATP mol−1 monomer min−1. As shown in figure 1, the flow dialysis demonstrates that TyrR binds to ATP with a half saturation value of 3.1 m. TyrR-(188-467) binds to ATP with a half saturation value of 7.6 m. TyrR-(188-467) had a 1.4 m dissociation constant when it coupled to rhodamine-ATP. These findings demonstrate that TyrR-(188-467) had a 2- to 5-fold reduced affinity for binding ATP and rhodamine-ATP than TyrR. Dixon, M. P., Pau, R. N., Howlett, G. J., Dunstan, D. E., Sawyer, W. H., & Davidson, B. E. (2002). The central domain of Escherichia coli TyrR is responsible for hexamerization associated with tyrosine-mediated repression of gene expression. Journal of Biological Chemistry, 277(26), 23186-23192.‏

Figure 1. Flow dialysis analysis of ATP binding to TyrR and TyrR-(188–467). The fractional saturation with ATP of 40 μM TyrR (closed circles) and TyrR-(188–467) (open circles) at various concentrations of ATP

References

1. Dixon, M. P., Pau, R. N., Howlett, G. J., Dunstan, D. E., Sawyer, W. H., & Davidson, B. E. (2002). The central domain of Escherichia coli TyrR is responsible for hexamerization associated with tyrosine-mediated repression of gene expression. Journal of Biological Chemistry, 277(26), 23186-23192.‏ Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 720
  Illegal BglII site found at 1062
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 526
  
• 1000
  COMPATIBLE WITH RFC[1000]