BBa_K218000 1 BBa_K218000 LuxPQ from Vibrio harveyi 2009-10-15T11:00:00Z 2015-05-08T01:11:30Z GenBank Accession: NC_009784 Vibrio harveyi ATCC BAA-1116 chromosome II, REGION: 579654..580751. Neiditch MB, Federle MJ, Pompeani AJ, Kelly RC, Swem DL, Jeffrey PD, Bassler BL, Hughson FM. Ligand-induced asymmetry in histidine sensor kinase complex regulates quorum sensing. Cell. 2006 Sep 22;126(6):1095-108. LuxP is a ABC-type sugar transport system periplasmic protein (Protein ID = YP_001447483.1). LuxQ is a histidine kinase (Protein ID = YP_001447484.1). Usage and Biology 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 (K131015); however, this part was cloned again and sequenced and is being re-submitted. LuxP and LuxQ are found in the operon, thus we decided to keep them together. We kept natural RBS in front of LuxP, as well as the intrinsic terminator. Silent mutations were performed on a EcoRI site located within LuxP, EcorI site within LuxQ, and XbaI site within LuxQ. false Jeremy Kubik annotation2048321 1 stop for LuxQ range2048321 1 3704 3706 annotation2048337 1 Silent mutation to remove XbaI site range2048337 1 1618 1618 annotation2048320 1 LuxP range2048320 1 30 1127 annotation2048336 1 Silent mutation to remove EcoRI site range2048336 1 1465 1465 annotation2048322 1 LuxQ range2048322 1 1127 3706 annotation2048335 1 Silent mutation to remove EcoRI site range2048335 1 599 599 annotation2048319 1 stop for LuxP range2048319 1 1125 1127 annotation2048299 1 start of LuxQ range2048299 1 1127 1129 annotation2048298 1 start of LuxP range2048298 1 30 32 BBa_K218000_sequence 1 atgctcgataaaaactaaaagagcaatagatgaagaaagcgttactattttccctgatttctatggtcggtttttctccagcgtctcaagcaacacaagttttgaatgggtactggggttatcaagagtttttggacgagtttcccgagcaacgaaatctgaccaatgctttatcagaagcagtacgagcacagccggtcccactgtctaaaccgacacaacgcccgattaaaatatcagtggtttacccaggacagcaagtttcagattactgggtaagaaatattgcatcattcgaaaaacgtttgtataagttgaatattaactaccaactgaaccaagtgtttactcgtccaaatgctgatatcaagcaacaaagcttgtcattaatggaagcgctcaagagcaaatcggattacttgattttcacgcttgatacgacaagacaccgtaaatttgttgagcacgttttggactcaacgaacaccaaattgatcttgcaaaatatcactacaccagtccgtgagtgggacaaacatcaaccgtttttatatgtcggatttgaccacgcagaaggcagtcgtgaattagcaacagagttcggaaagttcttcccaaaacacacatattacagtgtgctctacttttctgaaggttatattagcgatgtgagaggtgatacttttattcaccaagtaaaccgtgataataactttgagctacaatcagcgtattacacgaaggcaaccaagcaatccggctatgatgctgcgaaagcgagtttagcaaaacatccagatgttgattttatctatgcatgttcgaccgacgtagcattaggtgcagtagacgcactggctgagttgggacgtgaagatattatgatcaatggctggggtggaggctctgctgagttagacgctatccagaagggtgatttagacatcaccgtcatgcgtatgaatgatgacactggcatagccatggcagaagcgattaagtgggacttggaagataaaccagttccgaccgtatactcaggtgactttgaaatcgtaacaaaggcagattcaccggagagaatcgaagcgctgaaaaagcgcgcatttagatattcagataattgatgacaacaacgcgatcaaacattaaaaagcgtcgctcgctggcgacgctcataacaaagatcatcattttagttcttgccccaattattctggggattttcattcagagctattacttctccaagcaaatcatttggcaagaagtagaccgaaccaaacagcaaacctctgcactgatccacaatatatttgatagccactttgcggcgatccagatacatcatgacagtaattccaagagcgaagtcattcgtgatttctacactgatcgcgacacggatgtgctcaactttttcttcctcagtatcgaccaaagcgatccgtcgcacacaccagagttccgttttctaacggaccacaaaggcatcatttgggacgatggaaatgcgcatttctatggtgtgaacgaccttatccttgatagccttgccaatcgggtcagtttcagtaacaactggtattacattaatgtcatgacctccattggttcgagacacatgctcgtgcgccgtgtgccgatcctagacccttcaacaggagaggtgcttggtttctcatttaatgccgtcgtcttagacaacaacttcgctttgatggaaaagctcaagagtgaaagtaacgtcgacaatgtggtgctggttgctaatagcgttcctttagcaaactctttgattggtgatgagccatataacgttgctgatgtattgcagcgtaaaagttcagacaaaagactcgataagctgttggtaatagaaacgccaatcgtcgtaaatgcagtgactaccgagctttgcttgttgacggtacaagacaatcagagtgtggtgacattacaaatccaacatattctagccatgcttgcatcgatcatcggtatgatcatgattgccttaatgagtagggaatggattgagagtaaagtttcggcgcagttagaatctttgatgtcttacacccgctctgctcgtgaggaaaaagggtttgaacgatttggcggttcggatattgaagagtttgatcacatcggttcaacccttgaaagtacattcgaagagcttgaagcgcagaagaagtcgttccgagatctgtttaattttgccttatcacccatcatggtttggtctgaagagagtgtcctgattcagatgaaccctgccgcgcgcaaagaattagtgatcgaagacgatcatgaaatcatgcatccggtcttccaaggctttaaagagaaattgaccccacacctcaaaatggcggctcaaggtgcgacgttgactggggtgaacgtgcctattggtaataagatctaccgatggaacttgtcgccaattcgtgttgatggcgatatcagtggcattattgtgcaaggccaagacattacaacacttatcgaagccgagaagcagagtaacattgcgcgtagagaagcagaaaaatcggcgcaagcacgtgcagacttccttgctaaaatgagccatgaaattcgtacgccaatcaacggcattttaggtgtcgcccaattattgaaagattctgtcgatacacaagagcagaagaatcaaatcgacgtcctgtgccacagtggcgagcacttgcttgcagtactgaacgatattctcgatttctcaaagatagagcagggcaagttcaatattcagaaacacccgttctccttcaccgataccatgcgtacattggaaaatatttatcgtccgatttgcacaaataagggggtggagttggtcatcgagaatgagcttgacccgaatgttgaaatcttcaccgatcaagtccgcttgaatcagattctatttaacttagtgagtaatgccgttaagttcacgccgattggctcgattcgactgcacgcagaacttgaacaattctatggtgcggagaacagcgtgttagttgtggaactgactgatactggcatcggcattgaaagcgataagctcgaccaaatgttcgaaccttttgtgcaagaagagtcgacaaccacacgcgaatatggcggtagcggcctaggtttgaccatcgttaagaacctagtcgatatgttagaaggtgatgttcaggtccgcagtagcaaggggggggggacaacatttgttataacacttccagtaaaagatcgtgagcgtgtcttaaggcctctggaggtcagtcaacgtatcaagccggaagccttgtttgatgaaagtttaaaagtgctactggtggaagataaccataccaatgcgtttatccttcaggctttctgtaagaagtataaaatgcaggtggattgggcgaaagatgggctggacgcgatggagctcctttctgataccacctacgatctgatcctcatggataaccaattaccccaccttggtggtattgagaccacgcacgagattcgccagaacttgaggcttggaacgccaatttacgcgtgtacagcagacaccgcgaaagaaaccagtgatgcgtttatggcggcaggtgcaaactatgtcatgctgaagccaattaaagagaatgcgttacatgaggcgtttgtcgatttcaaacaacgtttcttggtagaaagaacctaacggtttaatggcagtgataagttaggggctgaagttaaataataaaagtaaagaagggagcgtaatgcactaatgccgttcacttaaggtgatcggcattgttttttctagggtatcgg 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