BBa_K218001 1 BBa_K218001 LuxOU from Vibrio harveyi 2009-10-16T11:00:00Z 2015-05-08T01:11:30Z GenBank Accession# L26221 LuxO Protein ID# AAD12736.1 LuxU Protein ID# AAD12737.2 Jeremy A. Freeman and Bonnie L. Bassler. Sequence and function of LuxU: a two-component phosphorelay protein that regulates quorum sensing in Vibrio harveyi. 1999. Journal of Bacteriology. Feb 1999, p. 899-906. Vibrio harveyi luminescence regulatory protein LuxO (GenBank accession# P0C5S5) and phosphorelay protein LuxU (GenBank accession# AAD12737). Quorum-sensing bacteria produce and release chemical signal molecules termed autoinducers (AIs) whose external concentration increases as a function of increasing cell-population density. Bacteria detect the accumulation of a minimal threshold stimulatory concentration of these autoinducers and alter gene expression, and therefore their behavior. Using these signal-response systems, bacteria synchronize particular behaviors on a population-wide scale and thus function as multicellular organisms. The bioluminescent marine bacterium Vibrio harveyi uses three different AIs???AHL, CAI-1, and AI-2???to control the expression of genes responsible for bioluminescence and numerous other traits. We have designed our System 2 based on V. harveyi AI-2 signaling. V. harveyi AI-2 signal is a furanosyl borate diester, production of which requires the LuxS enzyme. Biosynthesis of AI-2 is dependent on the usage of S-adenosylmethionine (SAM) by the cell in various methylation reactions. For this reason, during periods of exponential growth, there is a very large production of AI-2, thus perhaps signaling to neighbors that a suitable environment for growth (i.e. rich in nutrients) has been found. LuxS catalyzes the formation of the (S)-4,5-dihydroxy-2,3-pentanedione (DPD) intermediate which spontaneously cyclizes and reacts with borate to give AI-2. AI-2 is bound in the periplasm by the protein LuxP, which is constitutively bound to LuxQ, a membrane bound histidine kinase sensor. The binding of AI-2 to LuxP is necessary in regulating the activity of the periplasm-bound LuxQ. At low cell density, in the absence of significant amounts of autoinducers, LuxQ acts as a kinase, autophosphorylates, and subsequently transfers the phosphate to the cytoplasmic protein LuxU. LuxU passes the phosphate to the DNA-binding response regulator protein LuxO. Phospho-LuxO, in conjunction with a transcription factor termed σ54, involved in nitrogen metabolism, activates transcription of the genes encoding five regulatory small RNAs (sRNAs) termed Qrr1???5 (for Quorum Regulatory RNA). The Qrr sRNAs interact with an RNA chaperone termed Hfq, involved in mRNA splicing. The sRNAs, together with Hfq, bind to and destabilize the mRNA encoding the transcriptional activator termed LuxR. LuxR is required to activate transcription of the luciferase operon: luxCDABE. Thus, at low cell density, because the luxR mRNA is degraded, the bacteria do not express the genes necessary for bioluminescence. At high cell density, when the autoinducers accumulate to the level required for detection, the kinase activity of LuxQ is overtaken by its phosphatase activity and thus drains phosphate from LuxO via LuxU. Unphosphorylated LuxO cannot induce expression of the sRNAs. This allows translation of luxR mRNA, production of LuxR, resulting in bioluminescence. Reference: Waters C.M. and Bassler B.L. Quorum sensing: cell-to-cell communication in bacteria. Annu Rev Cell Dev Biol. 2005;21:319-46. false false _321_ 0 4333 9 It's complicated false This part was submitted to the Registry in 2008 (K131016); however, this part was cloned again and sequenced and is being re-submitted. LuxO and LuxU are found within the operon, therefore we have decided to keep these genes together. The internal RBS was also kept. false Jeremy Kubik annotation2048771 1 Stop for LuxO range2048771 1 1390 1392 annotation2048773 1 LuxO range2048773 1 31 1392 annotation2048775 1 Stop for LuxU range2048775 1 1731 1733 annotation2048770 1 Start of LuxO range2048770 1 31 33 annotation2048776 1 LuxU range2048776 1 1389 1733 annotation2048769 1 RBS range2048769 1 5 12 annotation2048774 1 Start of LuxU range2048774 1 1389 1391 BBa_K218001_sequence 1 acagaaggtcaaaagtctcgttatctacttatggtagaagacaccgcatccgttgcggcactttaccgctcttacctcacgccacttggcatcgatatcaatattgttggaacaggcagagacgccattgaaagcctgaaccatcgcattcctgatcttattctgctcgatcttcgtctacctgatatgacggggatggacgtattgcacgcggtgaagaaaagccacccagacgtgccaatcatcttcatgacagcccatggttctatcgatactgcggtagaggcgatgcgccacggttctcaagacttcctaatcaaaccatgtgaagcagaccgtttacgtgtcacggtgaacaatgcgatccgtaaagcaaccaaattaaagaatgaagctgacaaccccggtaaccaaaattaccaaggcttcatcggcagtagccaaacgatgcagcaggtttaccgcaccattgactcggcagcgagcagtaaagcgagtattttcatcacgggtgaaagtggtacgggtaaagaagtgtgtgccgaagcgattcacgcagcaagcaaacgcggtgataagccgtttatcgccatcaactgtgcggcaatcccgaaagaccttattgaaagtgagctgtttggtcacgtaaaaggtgcgtttactggtgctgcgaatgaccgacaaggtgcggcagagcttgctgatggcggcaccttgttccttgatgaactctgtgaaatggacttggatcttcaaactaagctattgcgctttatccaaacgggtacattccaaaaagtcggttcttctaaaatgaagagcgtggatgtgcgctttgtgtgtgcaactaaccgagacccttggaaagaagtgcaagaaggccgtttccgtgaagacttgtattaccgtttgtacgtgattcctttgcaccttccgccgctgcgtgagcgtggtaaagacgttattgaaattgcatactcgttgcttggttatatgtctcatgaggaaggtaagagtttcgtccgtttcgcacaagacgtgattgaaagattcaacagctacgaatggccgggtaacgttcgccagttgcaaaacgtattgcgtaatatcgtggtactgaacaatggcaaagagatcacgctggatatgttaccgccaccactgaatcagcctgttgtgcgccaatcggtagcaaaattcattgaacctgacattatgacggtgtcagatattatgccgctttggatgacagagaaaatggctattgagcaggcaattcaagcgtgtgaaggcaacattccacgcgctgctggctatttggatgttagtccatcaacgatttatcgcaagttgcaagcttggaatagcaaggacgaaaaacaaaacgtatgaatacggacgtattaaatcagcaaaaaattgaagaactgtctgcggaaattggtagcgataatgttcctgttttgcttgatatttttcttggggaaatggactcctacattggcactttaactgaacttcagggctcagagcagctgttgtatttaaaagagatcagccacgcactgaaaagtagtgctgccagctttggcgcagatcgattgtgtgaacgagcgattgccatcgacaagaaagcaaaagcgaatcaattgcaagagcaggggatggagacgagcgaaatgctcgctttacttcatatcactcgtgacgcctaccgttcttggacaaactaacgtttcgagcaagacattaagcgcaacgtaaaaacacaaagcccttccggtgtggaagggcttttttgtttggggagtttgctccgaatcgaagccgctttctcaatgcttttcgtctagttagacagtaagcgctccataaaacccgcattctaatcgcctagcgcgaagaataagatcaagtctccaaccatgaggagatttgaaatggg igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 James Alastair McLaughlin Chris J. Myers 2017-03-06T15:00:00.000Z