Part:BBa_K4375004
BLADE (light-responsive AraC transcription factor)
BLADE is a modified AraC transcription factor engineered to react to blue light instead of arabinose. The AraC’s arabinose binding domain was swapped to a homodimerizing Vivid domain. Upon blue light illumination, the protein dimerizes and the AraBAD promoter becomes active.
Usage and Biology
BLADE protein is produced from AraC by exchanging the dimerization domain of AraC with a light-inducible dimerization domain. The switching of this engineered AraC from monomer to dimer is controlled with light. In its monomeric form, the engineered AraC would contact the high-affinity I1 half-site16, but not the low-affinity I2 half-site, needed to recruit the RNA polymerase. For the light-triggered dimerization domain Vivid (VVD) domain was selected, which has often been successfully used to control, with light, the dimerization of proteins of interest. VVD senses blue light via the flavin adenine dinucleotide (FAD) chromophore. Blue light triggers the formation of a cysteinyl-flavin adduct, which generates a new hydrogen-bond network that releases the N terminus (N-terminal cap) from the protein core and restructures it, creating a new dimerization interface.
Characteristics
For experimental results with this part please refer to BBa_K4375019 and BBa_K4375003.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
References
Romano, E.; Baumschlager, A.; Akmeriç, E. B.; Palanisamy, N.; Houmani, M.; Schmidt, G.; Öztürk, M. A.; Ernst, L.; Khammash, M.; Di Ventura, B. Engineering AraC to Make It Responsive to Light Instead of Arabinose. Nature Chemical Biology 2021, 17 (7), 817–827. https://doi.org/10.1038/s41589-021-00787-6.
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