Vibrio Fischeri:

1. Vibrio Fischeri: Bioluminescence and Quorum Sensing Jacob Northey FW4089 Project

2. Bioluminescence • Has evolved separately many times • Usually involved in symbiotic or cooperative relationships • Fireflies lighting at the same time • Squids and light organs • Odd bioluminescence • Night vision (Aristostomias)Aristostomias

3. How it works • Chemicals Involved • Luciferase: Enzyme that catalyzes reaction • Luciferin: Chemical that produces light • Reaction occurs by the oxidation of luciferin • Products are light and an oxyluciferin • The luciferin in V. fischeri is FMNH2 picture above

4. Vibrio Fischeri: Symbiotic • When existing as a population in the stomach of a squid, V.fischeri emit light • Light is the color of the moon • This acts as a camoflague to the squid at night • In return, V.fischeri gets a competition free home to live in • Yet when the bacterium exist in less dense populations they don’t emit light

5. Quorum Sensing • If a V. fischeri is not densely populated then it must not be inside a host • It is inefficient to produce light • All V. fischeri produce a protein called an autoinducer • The autoinducer is used as a transcription factor • More autoinducer means more light

6. Quorum Sensing (cont.) • VAI (Vibrio AutoInducer) is produced by the luxI gene • VAI binds to a protein produced by luxR • This activates the luxR which is a transcription activator binding to the luxP operon • This begins transcription of the luciferase genes

7. Lux Gene System LISTENING IN ON BACTERIA: ACYL-HOMOSERINE LACTONE SIGNALLING. Nature Reviews Molecular Cell Biology3; 685-695 (2002); doi:10.1038/nrm907

8. 1 gtcgaccttc ctggttcaga gcctcatatc catattacga ccacttcaat gggcgatcaa 61 aaagtacatt acatgatttt ttgcccaaca gaaaaagcct ctcatttaga gcagcttatt 121 cgtcaagatt tcatggaaat gtatgagatc aattttccac aaaaaacaga gtaacaaaga 181 gaaaaaccta taggtataaa gctttactta cgtacttaat ttttaaagta tgggcaatca 241 attgctcctg ttaaaattgc tttagaaata ctttggcagc ggtttgttgt attgagtttc 301 atttgcgcat tggttaaatg gaaagtgaca gtacgctcac tgcagcctaa tatttttgaa 361 atatcccaag agctttttcc ttcgcatgcc cacgctaaac attctttttc tcttttggtt 421 aaatcgttgt ttgatttatt atttgctata tttatttttc gataattatc aactagagaa 481 ggaacaatta atggtatgtt catacacgca tgtaaaaata aactatctat atagttgtct 541 ttttctgaat gtgcaaaact aagcattccg aagccattgt tagccgtatg aatagggaaa 601 ctaaacccag tgataagacc tgatgttttc gcttctttaa ttacatttgg agatttttta 661 tttacagcat tgttttcaaa tatattccaa ttaattggtg aatgattgga gttagaataa 721 tctactatag gatcatattt tattaaatta gcgtcatcat aatattgcct ccatttttta 781 gggtaattat ctagaattga aatatcagat ttaaccatag aatgaggata aatgatcgcg 841 agtaaataat attcacaatg taccatttta gtcatatcag ataagcattg attaatatca 901 ttattgcttc tacaagcttt aattttatta attattctgt atgtgtcgtc ggcatttatg 961 tttttcatac ccatctcttt atccttacct attgtttgtc gcaagttttg cgtgttatat 1021 atcattaaaa cggtaatgga ttgacatttg attctaataa attggatttt tgtcacacta 1081 ttgtatcgct gggaatacaa ttacttaaca taagcacctg taggatcgta caggtttacg 1141 caagaaaatg gtttgttata gtcgaataaa cgcaagggag gttggtatga ctataatgat 1201 aaaaaaatcg gattttttgg caattccatc ggaggagtat aaaggtattc taagtcttcg 1261 ttatcaagtg tttaagcaaa gacttgagtg ggacttagtt gtagaaaata accttgaatc 1321 agatgagtat gataactcaa atgcagaata tatttatgct tgtgatgata ctgaaaatgt 1381 aagtggatgc tggcgtttat tacctacaac aggtgattat atgctgaaaa gtgtttttcc 1441 tgaattgctt ggtcaacaga gtgctcccaa agatcctaat atagtcgaat taagtcgttt 1501 tgctgtaggt aaaaatagct caaagataaa taactctgct agtgaaatta caatgaaact 1561 atttgaagct atatataaac acgctgttag tcaaggtatt acagaatatg taacagtaac 1621 atcaacagca atagagcgat ttttaaagcg tattaaagtt ccttgtcatc gtattggaga 1681 caaagaaatt catgtattag gtgatactaa atcggttgta ttgtctatgc ctattaatga 1741 acagtttaaa aaagcagtct taaattaatg ttgttaaatc attaatttat tttaaatact

9. 1801 aagtatatta tagggaaaat aatgaataaa tgtattccaa tgataattaa tggaatgatt 1861 caagattttg ataattatgc atataaagaa gttaaactaa ataatgataa tagagtaaaa 1921 ttatctgtca ttactgaaag ttcagtttca aaaacattaa atatcaaaga tagaattaat 1981 ctaaatttaa atcagattgt gaatttttta tataccgttg gtcaacgatg gaaaagtgaa 2041 gaatataatc ggcgacgaac ctatatccgt gagttaaaaa catatcttgg ttattctgat 2101 gaaatggcaa gattagaagc gaattggatt gcaatgttat tgtgctctaa aagtgctttg 2161 tatgacattg ttaattatga tttgggctct atacacgtat tagatgaatg gcttccacgt 2221 ggtgattgct atgttaaagc acaaccgaaa ggtgtttctg ttcacttgtt agctggtaat 2281 gttccattat caggagtgac atctattttg cgtgctattt taacaaaaaa tgagtgcatt 2341 attaaaactt cgtcttcaga tccttttact gcaaacgctt tagtttccag ttttattgat 2401 gttaatgcag accatccaat aaccaaatca atgtctgtta tgtattggcc gcatgatgaa 2461 gatatgactc tatctcaaag aataatgaat catgccgacg tggttattgc ttggggtgga 2521 gacgaggcga ttaaatgggc ggtaaaatat tcaccaccgc atgtcgatat tctgaaattt 2581 ggaccaaaga aaagcttaag tattattgaa gctcctaaag atatagaagc cgcagcaatg 2641 ggggttgctc atgatatttg tttctatgac cagcaagcct gcttctctac tcaagacgtt 2701 tattatatag gagataattt acctttattt ttaaatgaac ttgaaaaaca gctagatcga 2761 tacgcgaaaa ttttaccaaa aggttcaaat agttttgatg aaaaagcggc gtttactctt 2821 actgaaaaag aaagtctatt tgctggatat gaagtgagaa agggagataa gcaagcttgg 2881 ttaatagtcg tatcacctac aaatagcttt ggaaatcaac cgctatcacg aagtgtgtat 2941 gttcatcaag tatctgatat taaagagata attccttttg ttaataaaaa tagaacacaa 3001 actgtttcta tttatccttg ggaagcgtca ttaaaatatc gagataaatt agcaagaagt 3061 ggagttgaaa gaattgttga atcaggcatg aataatattt tcagagttgg aggggctcat 3121 gattcattat ctcctctcca gtacctagtt aggtttgtat cgcatgagag accatttaat 3181 tatacgacaa aagatgttgc ggttgaaatc gaacaaacac gttacttaga ggaagataag 3241 tttttagttt ttgtcccata gttaaaggaa attatatgaa agatgaaagt gcttttttta 3301 cgattgatca catcatcaag cttgataatg gtcagtctat ccgagtttgg gaaacactcc 3361 ctaaaaagaa cgtaccagag aaaaaacata caatacttat tgcttcgggt tttgctagaa 3421 gaatggatca ttttgcaggt cttgctgagt atttatctac taacggtttt catgtcattc 3481 gctacgattc tttgcatcat gttggattaa gcagtggatg tataaatgaa tttacgatgt 3541 cgattggaaa aaatagcctg cttacagtcg tagattggct tacagatcat ggtgtcgaac 3601 gaatagggct gattgctgct agtttgtcag cgagaatcgc ctatgaggta gtaaataaaa 3661 ttaaattatc atttttaatt acggccgtag gtgtcgttaa tcttagagat acattagaaa 3721 aagcattgga gtatgactat ttgcaattac ctatttcaga gttaccagaa gatcttgact 3781 ttgaaggtca taatttagga tcggaggtct ttgttacaga ttgctttaaa cataattggg 3841 acacgttaga ctcgacactt aatagtgtta aaggattagc gattccattt attgctttta 3901 ctgcaaacga tgatagttgg gtaaagcaaa gtgaagttat agagctcatt gatagcattg 3961 aatctagtaa ttgtaagctc tattcgctaa ttggaagttc acatgatctt ggggaaaatt 4021 tggttgtatt aagaaatttt tatcaatcag tgacgaaggc agccttagca ttagatgatg 4081 gtttattgga tttagagata gacattattg aacctcgatt tgaggacgtt acaagtatta 4141 ctgttaagga gcgtagatta aaaaatgaaa ttgaaaatga attattagaa ttggcttaaa

10. 4201 taaacagaat caccaaaaag gaatagagta tgaagtttgg aaatatttgt ttttcgtatc 4261 aaccaccagg tgaaactcat aagcaagtaa tggatcgctt tgttcggctt ggtatcgcct 4321 cagaagaggt agggtttgat acatattgga ccttagaaca tcattttaca gagtttggtc 4381 ttacgggaaa tttatttgtt gctgcggcta acctgttagg aagaactaaa acattaaatg 4441 ttggcactat gggggttgtt attccgacag cacacccagt tcgacagtta gaagacgttt 4501 tattattaga tcaaatgtcg aaaggtcgtt ttaattttgg aaccgttcga gggctatacc 4561 ataaagattt tcgagtattt ggtgttgata tggaagagtc tcgagcaatt actcaaaatt 4621 tctaccagat gataatggaa agcttacaga caggaaccat tagctctgat agtgattaca 4681 ttcaatttcc taaggttgat gtatatccca aagtgtactc aaaaaatgta ccaacctgta 4741 tgactgctga gtccgcaagt acgacagaat ggctagcaat acaagggcta ccaatggttc 4801 ttagttggat tattggtact aatgaaaaaa aagcacagat ggaactctat aatgaaattg 4861 cgacagaata tggtcatgat atatctaaaa tagatcattg tatgacttat atttgttctg 4921 ttgatgatga tgcacaaaag gcgcaagatg tttgtcggga gtttctgaaa aattggtatg 4981 actcatatgt aaatgcgacc aatatcttta atgatagcaa tcaaactcgt ggttatgatt 5041 atcataaagg tcaatggcgt gattttgttt tacaaggaca tacaaacacc aatcgacgtg 5101 ttgattatag caatggtatt aaccctgtag gcactcctga gcagtgtatt gaaatcattc 5161 aacgtgatat tgatgcaacg ggtattacaa acattacatg cggatttgaa gctaatggaa 5221 ctgaagatga aataattgct tccatgcgac gctttatgac acaagtcgct cctttcttaa 5281 aagaacctaa ataaattact tatttgatac tagagataat aaggaacaag ttatgaaatt 5341 tggattattt tttctaaact ttcagaaaga tggaataaca tctgaagaaa cgttggataa 5401 tatggtaaag actgtcacgt taattgattc aactaaatat cattttaata ctgcctttgt 5461 taatgaacat cacttttcaa aaaatggtat tgttggagca cctattaccg cagctggttt 5521 tttattaggg ttaacaaata aattacatat tggttcatta aatcaagtaa ttaccaccca 5581 tcaccctgta cgtgtagcag aagaagccag tttattagat caaatgtcag agggacgctt 5641 cattcttggt tttagtgact gcgaaagtga tttcgaaatg gaatttttta gacgtcatat 5701 ctcatcaagg caacaacaat ttgaagcatg ctatgaaata attaatgacg cattaactac 5761 aggttattgt catccccaaa acgactttta tgattttcca aaggtttcaa ttaatccaca 5821 ctgttacagt gagaatggac ctaagcaata tgtatccgct acatcaaaag aagtcgtcat 5881 gtgggcagcg aaaaaggcac tgcctttaac atttaagtgg gaggataatt tagaaaccaa 5941 agaacgctat gcaattctat ataataaaac agcacaacaa tatggtattg atatttcgga 6001 tgttgatcat caattaactg taattgcgaa cttaaatgct gatagaagta cggctcaaga 6061 agaagtgaga gaatacttaa aagactatat cactgaaact taccctcaaa tggacagaga 6121 tgaaaaaatt aactgcatta ttgaagagaa tgcagttggg tctcatgatg actattatga 6181 atcgacaaaa ttagcagtgg aaaaaacagg gtctaaaaat attttattat cctttgaatc 6241 aatgtccgat attaaagatg taaaagatat tattgatatg ttgaaccaaa aaatcgaaat 6301 gaatttacca taataaaatt aaaggcaatt tctatattag attgcctttt taaatttctg

11. 6361 ttgatattag gtattactgg agagggtatg actgtccata ctgaatataa aagaaatcaa 6421 atcattgcta gttcagaaat tgatgatctt atctttatga cgaaaccaca agagtggtca 6481 tttgaagagc aaaaagaaat acgggataaa ttagttcgtg aggcttttta ttttcactac 6541 aatagaaatg aagaatatag aaattattgt atcaatcagc atgtgagtga taatttacac 6601 actattgatg aaatacccgt gtttccaaca tctgttttta aatataagaa attacatact 6661 gtcacagccg aggacattga aaattggtat acaagtagtg gaactcgtgg agtaaaaagt 6721 catattgcac gtgatcgtct tagcattgaa cgcttgcttg gttctgtcaa cttcggaatg 6781 aaatacgttg gagattggtt tgagcatcaa atggaattga taaatttagg accagataga 6841 ttcaatacaa ataatatttg gtttaaatat gtcatgagtt tggtcgagtt actttatccg 6901 actgaattta cagttgataa tgacaaaata gattttgaaa aaacagtaaa acatctattt 6961 agaattaaga atagtaaaaa agacatttgc ttaattgggc caccattttt tgtgtatctt 7021 ttgtgccaat atatgaaaga aaacaatatt gaatttaaag gaggagatag agtacatatt 7081 attactggtg gaggatggaa atctaatcag aatgactctt tagatcgtgc tgattttaat 7141 caattattaa tggatacttt ccaactcgac aagattaatc aaattagaga tacctttaat 7201 caagttgagc ttaatacttg tttttttgaa gatgaatttc aaagaaaaca tgttccaccg 7261 tgggtatatg ctcgggctct tgatcctgaa accttgaaac ccgtagcaga tggtgagatc 7321 gggttgttaa gttatatgga cgcctcatca actgcttacc ctgcttttat tgttactgat 7381 gatatcggta ttgtaaaaga aattagagaa ccagatcctt acccaggggt aactgttgag 7441 attgttcggc gcttaaatac acgtgcgcaa aaaggatgcg cgctctctat ggctaatgtc 7501 atacaaaaga atatcaagga ttaagttatg attgttgatg gtagagtttc aaaaatagta 7561 ttagcgtcaa taagaaataa tatatataag gtatttatta ctgtaaattc accaataaag 7621 ttcatcgctg gacaatttgt aatggtcacg attaatggga aaaaatgccc tttttcaatt 7681 gcgaattgcc cgacaaaaaa ttacgaaata gaattgcata ttggtagttc gaatagagac 7741 tgctcattgg atattatcga atattttgtc gatgctcttg ttgaggaagt cgcaattgag 7801 ttagatgctc cccatggaaa cgcttggtta cggtctgaaa gtaataaccc attgctatta 7861 attgcgggag gtacaggttt atcatatata aatagcattc taacaaattg cttaaatagg 7921 aatatacctc aagatattta tctttactgg ggagtaaaag acagttctct tttgtatgaa 7981 gatgaagagt tactaaactt atcactaaac aacaaaaact ttcattatat tcctgttatt 8041 gaagataaaa gtgaagaatg gatagggaga aaaggcactg ttcttgatgc tgtcatggaa 8101 gattttactg atcttactta ttttgatatt tatgtttgtg gacccttcat gatggctaaa 8161 acagcaaaag aaaaattaat tgaagagaaa aaagcaaagt cagaacagat gtttgccgat 8221 gcttttgcat acgtataaag agaatataaa aagccagatt attaatccgg cttttttatt 8281 atttcaatgt gtccaagggt aaccaagtta cttcaactcc cgcctgagag aacatatctt 8341 ggctcacttg aattttttca ccccaacgag ataagaagtc ttcagtttgt tcagggcaat 8401 atacttttga tattcctgtt tggatgattt ttgctgcgca gtttggacaa gggaagtgag 8461 taacccaaat atcgcaacct tctaaatcac gcttggcaaa taaaatggca ttttcttcag 8521 catgaagtgt ttttaaatat ttaatttcac gttcatctgt atctgcactg tctgatacac 8581 catgtgggta gccattaaat ccaacagaaa caatacggtt atgcttggtg attactgcac 8641 caacttgagt cgac

12. Lux Autoinducer LISTENING IN ON BACTERIA: ACYL-HOMOSERINE LACTONE SIGNALLING. Nature Reviews Molecular Cell Biology3; 685-695 (2002); doi:10.1038/nrm907

13. Autoinducer BLAST

14. LuxR Promoter • luxR binding to the luxP binding site • luxR must be activated by the autoinducer VAI

15. References Bioluminescence Web Page. http://www.lifesci.ucsb.edu/~biolum/ Finney, Angela H. et al. “Role of the C-Terminal Domain of the Alpha Subunit of RNA Polymerase in LuxR-Dependent Transcriptional Activation of the lux Operon during Quorum Sensing” Journal of Bacteriology, 4520-4528 August 2002, 184 (16) Fuqua, Clay & Greenberg, E. Peter. “Listening in on Bacteria: Acyl-Homoserine Lactone Signalling.” Nature Reviews Molecular Cell Biology3; 685-695 (2002); doi:10.1038/nrm907