Part:BBa_K3610047
CORE ectodomain / mCherry N-terminal for S. cerevisiae
This part entails the ectodomain of the plant pattern recognition receptor COREfrom <S. lycopersicum</i> which is fused to the N-terminal domain of the mCherry protein via a 15 amino acid linker. The C-terminal sequence of the receptor protein, encoding the signal peptide, was removed from the sequence. To ensure localization, the secretion signal of the alpha Factor from yeast is added instead.
Usage and Biology
CORE
The cold shock protein receptor (CORE) is a plant pattern recognition receptor (PRR) and as such activates host innate immunity through detection of pathogen-associated molecular patterns (PAMPs). CORE is a leucine-rich repeat receptor-like kinase with 22 LRRs, there additionally is a 6 amino acid insert at LRR 11. It consists of an extracellular domain that perceives an epitope, csp22, from the highly conserved nucleic acid binding motif RNP-1 of bacterial cold-shock proteins (CSPs), which are highly abundant proteins found in the cytosol of bacteria. Further domains are a single pass transmembrane domain and an intracellular kinase domain (The sequence encoding the kinase domain is not in this part). Interaction of CORE with brassinosteroid-associated kinase (BAK)1 is necessary for inducing an immune response in the plant. The dimerization of CORE and BAK1 depends on the csp22, the ligand of CORE. The function of CORE in S. lycopersicum has been confirmed by expressing the receptor in A. thaliana, which made the plant responsive to csp22, a PAMP that is otherwise not perceived by PRRs from A. thaliana.
Usage with mCherry
In this case, the C-terminal domain of CORE, entailing the intracellular kinase domain, was removed from the sequence. Instead, the N-terminal domain of the split mCherry was fused to the C-terminal domain via a 15 amino acid linker.
The ligand-dependent interaction of CORE with its coreceptor BAK1 is driven by the extracellular ligand-binding domain. Further necessary is the transmembrane domain, including the juxtamembrane domain.
It is possible that dimerization of the two receptors can be induced without the intracellular kinase domain of neither CORE nor BAK1. We were interested in testing whether coexpression of this part with BAK1 fused to the N-terminal domain of split-mCherry in S. cerevisiae would allow to visually capture the presence of the bacterial epitope csp22 in water samples by ligand-dependent interaction of the two receptors, an interaction which would drive the reconstitution of the split-mCherry protein.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1740
Illegal BamHI site found at 373
Illegal BamHI site found at 1765
Illegal BamHI site found at 2117 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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