Team:Davidson-Missouri Western/Lsr

From 2008.igem.org

Lsr (AI-2) cell signaling system

Responsible: Kelly Davis, Xiao Zhu

Signal molecule: AI-2 (furanosyl borate diester in V. harveyi, a variety of other molecules in other species), all are derived from DPD. AI-2 is R-THMF in E. coli

[1]

Bacterial species:

lsrA,B,C,D,F,G,R,K: Escherichia coli HS, SMS-3-5, str. K12 substr. MG1655, and substr. DH10B.

lsrE:Escherichia coli str. K12 substr. MG1655

LuxS:Escherichia coli HS, SMS-3-5, APEC O1, str. K12 substr. MG1655, substr. DH10B, and UTI89.

Receiver protein: LsrR protein receives signal from sensor protein

Signal molecule synthase: Pfs enzyme, then LuxS autoinducer synthase

Target genes: lsr operon, including ABC transporter and LsrK kinase

Regulation: LsrR represses the lsr operon, derepression by phospho-AI-2; catabolite repression influences AI-2 accumulation through the cyclic AMP (cAMP)-cAMP receptor protein (CRP) complex, which directly stimulates transcription of the lsr operon and indirectly represses luxS expression.cAMP-CRP is shown to bind to a cAMP receptor protein (CRP) binding site located in the upstream region of the lsr promoter and works with the LsrR repressor to regulate AI-2 uptake.

Note: AI-2 is synthesized and secreted during exponential growth and is imported in stationary phase when glucose becomes limiting. In the presence of glucose, AI-2 is not imported because the lsr operon is not transcribed due to camp-CAP mediated repression. Both glycerol and G3P(glycerol 3-phosphate) repress lsr transcription, while the majority repression comes from G3P. DHAP represses lsr transcription by a cAMP-CAP-independent mechanism involving LsrR.

Parts Needed:

LsrR pro/term

LsrK

LsrACDB (transport)

LsrFGE (catabolic)

LuxS

Pfs enzyme (?)