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Presentation 10 mm Pontodrilus litoralis Ceder Key, Florida. 12 Aug 2014 Ceder Key, Florida. 12 Aug 2014 10 mm Pontodrilus litoralis Pontodrilus litoralis Evolution of firefly luciferase and Biosynthetic origin of firefly luciferin Yuichi Oba Graduate School of Bioagricultural Sciences, Nagoya University, Japan [email protected] 2014 Aug 13, 14:20-14:45 Firefly Symposium 2014 Gainesville, FL, USA ✓Bioluminescence mecahnism is common in all four coleopeteran families. Bioluminescence reaction in beetles 2+ N N C O O H ATP, Mg , O2 N N O + Light H O S S Beetle luciferase H O S S (> 46% AA similarity) Beetle luciferin Oxyluciferin Lampyridae Phengodidae Rhagophthalmidae Elateridae (Aquatica lateralis) (Phengodes sp.) (Rhagophthalmus ohbai) (Pyrophorus angustus) Photo of Phengodes sp.: http://www.flickriver.com/groups/1247869@N24/pool/interesting/ ✓Bioluminescence mecahnism is common in all four coleopeteran families. ✓Bioluminescence for each group was evolved independently. Bioluminescence reaction in beetles 2+ N N C O O H ATP, Mg , O2 N N O + Light H O S S Beetle luciferase H O S S (> 46% AA similarity) Beetle luciferin Oxyluciferin Luciferins in other arthropods O H O H O H N H H N O N N N a O 2C N H H N H N NH N H N NH 2 Lampyridae PhengodidaeH Rhagophthalmidae Elateridae N a O C O (Aquatica lateralis) (Phengodes sp.) (Rhagophthalmus ohbai) (Pyrophorus2 angustus) Ostracoda Krill (Vargula hilgendorfii) (MeganyctiphanesKrill (Crustacean) norvegica) What was the firefly luciferase evoleved from ? Firefly MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTD-AHIEVNITYAEYFEM Vargula -MKIIILSVILAYCVTVNCQDACPVEAEPPSSTPTVPTSCEAKEGECIDTRCATCKRDIL American firefly Photinus pyralis luciferase . :. * .:: * :. ** : .: *:.. : .: : Firefly SVRLAEAMK-RYGLNTNHRIVVCSENSLQFFMPVLGALFIG-----VAVAPANDIYNERE vs Vargula SDGLCENKPGKTCCRMCQYVIECRVEAAGYFRTFYGKRFNFQEPGKYVLARGTKGGDWSV * *.* : . : :: * :: :* .. * * .:* ... : ostracod Vargula hilgendorfii luciferase Firefly LLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSMYTFVTSHLP Vargula TLTMENLDGQKGAVLTKTTLEVAGDVIDITQATADPITVNGGADPVIANPFTIGEVTIAV *. *:. ..::*. *: :* 。 . * ::. :* :.: .: Firefly PGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHARDPIFGNQIIP Vargula VEIPGFNITVIEFFK----LIVIDILGGRSVRIAPDTANKGLISGICGNLEMNDADDFTT : :::. .* : :::: *. .: . .:: :. : .::: . Firefly DTAILSVVP-FHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKIQSALLVPTLF Vargula DADQLAIQPNINKEFDGCPFYGNPSD----IEYCKGLMEPYRAVCRNNINFYYYTLSCAF *: *:: * ::: *. * : : * : :: :. * Firefly SFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYGLTETTSAILIT Vargula AYCMGGEERAKHVLFDYVETCA--APETRGTCVLSGHTFYDTFDKARYQFQGPCKELLMA :: 。*: * : * .: ** :: 。.: *: . : * : . :*:: Firefly PEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSGYVNNPEATNALI Vargula ADCYWNTWDV-KVSHRDVESYTEVEKVTIRKQSTVVDLIVDGKQVKVGGVDVSIPYSSEN .: :. * ** . .:::. 。. :* * * : * *: . .: Firefly DKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILLQHP-----NIFDA Vargula TSIYWQDGDILTTAILPEALVVKFNFKQLLVVHIRDPFDGKTCGICGNYNQDSTDDFFDA . * ... :: * :.: .:*.*: : : .: .* :: ::*** Firefly G-VAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVFVDEVPKGL Vargula EGACALTPNPPGCTEEQKPEAERLCNSLFDSSIDEKCNVCYKPDRIAR--CMYEYCLRGQ ...*. : .* *: . .. :* .. ...:: : : :* Firefly TGKLDARKIREILIKAKKGGKSKL----- Vargula QGFCDHAWEFKKECYIKHGDTLEVPPECQ * * : *:*.. :: (no significant homology) Firefly MEDAKNIKKGPAPFYPLEDGTAGEQL---HKAMKRYALVPGTIAFTDAHIEVNITYAEYF American firefly Photinus pyralis luciferase Parsley MGDCVAPKEDLIFRSKLPDIYIPKHLPLHTYCFENISKVGDKSCLINGATGETFTYSQVE * *. *:。 * * ::* .::. : * .. .: :。.:**:: vs Firefly EMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPANDIYNERELLNS Parsley LLSRKVASGLNKLGIQQGDTIMLLLPNSPEYFFAFLGASYRGAISTMANPFFTSAEVIKQ Parsley 4-coumarate:CoA ligase :* ::*..::: *:: .. *:: ** ::*:..*** : *. : ** ::.. *:::. Firefly MNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSMYTFVTSHLPPGFN parsley LKASLAKLIITQACYVDKVKDYAAEK-NIQIICIDDAPQDCLHFSKLMEADESEMP---- :: * ..:::.. ::*: : : ** * * *: * *..: *.:* Firefly EYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHARDPIFGNQIIP-DTA parsley ------EVVIDSDDVVALPYSSGTTGLPKGVMLTHKGLVTSVAQQVDGDNPNLYMHSEDV * . .. :. : ***:******* *.*: . .:: * * : : . Firefly ILSVVPFHHGFGMFTTLGY-LICGFRVVLMYRFEEELFLRSLQDYKIQSALLVPTLFSFF parsley MICILPLFHIYSLNAVLCCGLRAGVTILIMQKFDIVPFLELIQKYKVTIGPFVPPIVLAI ::.::*:.* :.: :.* * .*. :::* :*: **. :*.**: . :**.:. : Firefly AKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYGLTETTSAILITPEG parsley AKSPVVDKYDLSSVRTVMSGAAPLGKELEDAVRAKFPNAKLGQGYGMTEAGPVLAMCLAF ***.::******.:: : **.***.**: :** :* . : ****:**: ..: : Firefly DDKP-----GAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSGYVNNPEATNA parsley AKEPYEIKSGACGTVVRNAEMKIVDPETNASLPRNQRGEICIRGDQIMKGYLNDPESTRT .:* ** *.** * *:** :*. :* *****:*:** **.**:*:**:*.: Firefly LIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILLQHPNIFDAGVA parsley TIDEEGWLHTGDIGFIDDDDELFIVDRLKEIIKYKGFQVAPAELEALLLTHPTISDAAVV **::****:***.: *:*:.:*******.:*****:********::** **.* **.*. Firefly GLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVFVDEVPKGLTGKL parsley PMIDEKAGEVPVAFVVRTNGFTTTEEEIKQFVSKQVVFYKRIFR-VFFVDAIPKSPSGKI : *:.***:*.*.** :* * **:** ::*:.**. *:: *.*** :**. :**: Firefly DARKIREILIKAKKGGKSKL parsley LRKDLRAKIASGDLPK---- :.:* : ... (34% amino acid identity) Schröder J (1989) Nucleic Acid Res. 17, 460. Homologous gene of Firefly MED---AKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAHIEVNITYAEYF CG6178 MTSKLLPGNIVYG-GPVTERQAQDSRSLGQYILDKYKSFGDRTVLVDAVNGVEYSASFMH beetle luciferase in * . ** * .*. : : . : :.:* . .:.** *: : : . Drosophila (CG6178) Firefly EMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPANDIYNERELLNS CG6178 KSIVRLAYILQKLGVKQNDVVGLSSENSVNFALAMFAGLAVGATVAPLNVTYSDREVDHA : **** ::: *:: *. : :.****::* :.::..* :*.:*** * *.:**: :: Firefly MNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSMYTFVTSHLPPGFN CG6178 INLSKPKIIFASKITIDRVAKVASKNKFVKGIIALSGTS--KKFKNIYDLKELMEDEKFK :*:*:*.::*.** :::: :* .* ::: ** :...: : *:.:* : *: Firefly -EYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHARDPIFGNQIIPDTA CG6178 TQPDFTSPAANKDEDVSLIVCSSGTTGLPKGVQLTQMNLLATLDSQIQPTV--IPMEEVT : **.. : ::*: ::**: ***:******* *.: . :. :* . : :.: Firefly ILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKIQSALLVPTLFSFFA CG6178 LLTVIPWFHAFGCLTLITTACVGARLVYLPKFEEKLFLSAIEKYRVMMAFMVPPLMVFLA :*:*:*:.*.** :* : * *:* : :***:*** :::.*:: *::**.*: *:* Firefly KSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYGLTETTSAILITPEGD CG6178 KHPIVDKYDLSSLMVLLCGAAPLSRETEDQIKERIGVPFIRQGYGLSESTLSVLVQNDEF * .::******.* : .*.****:*. : : :*: :* *******:*:* ::*: : Firefly DKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSGYVNNPEATNALIDKDG CG6178 CKPGSVGVLKVGIYAKVIDPDTGKLLGANERGELCFKGDGIMKGYIGDTKSTQTAI-KDG ***:** : : ***:* **** **.*:*****.:* **.**:.:.::*:: * *** Firefly WLHSGDLAYWDEDEHFFIVGRLKSLIKYKGYQVAPAELESILLQHPNIFDAGVAGLPDDD CG6178 WLHTGDIGYYDDDFEFFIVDRIKELIKYKGYQVPPAEIEALLLTNDKIKDAAVIGKPDEE ***:**:.*:*:* .****.*:*.*********.***:*::** : :* **.* * **:: Firefly AGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVFVDEVPKGLTGKRDARKIR CG6178 AGELPLAFVVKQANVQLTENEVIQFVNDNASPAKRLRGGVIFVDEIPKNPSGKILRRILR ***** *.** : . :**:*::::* .:.:.**:*****:****:**. :** * :* Firefly EILIKAKKGGKSKL CG6178 EMLKKQK----SKL *:* * * *** (41% amino acid identity) Molecular phylogenetic tree of beetle luciferases and the homologues (Bayesian tree) 1.00/99 0.1 changes Diaphanes pectinealis (DQ408300)* 0.97/99 Lampyris turkestanicus (AY742225) Nyctophila cf. caucasica (DQ072141)* 1.00/89 1.00/100 Lampyris noctiluca (X89479) 1.00/100 Pyrocoelia rufa (AF328553) 0.98/99 1.00/100 Pyrocoelia miyako (L39928) Lampyrinae Pyrocoelia pectoralis (EF155570)* 1.00/100 Pyrocoelia pygidialis (ACF61063) Cratomorphus distinctus (AY633557) Photinus pyralis (AB644228) Lucidina biplagiata (AB535101) 1.00/100 Luciola cruciata LcLuc1 (AB220162) 1.00/- 1.00/97 Aquatica lateralis LlLuc1 (AB644226) Luciola italica (DQ138966) 1.00/100 1.00/99 Luciola mingrelica (S61961) 0.56/64 Luciolinae 0.88/86 Luciola parvula (AB644227) 1.00/94 Luciola unmunsana (AF420006) 0.99/- 0.93/- 1.00/100 Lampyroidea maculata (DQ137139) Luciola terminalis (EU302126)* LAMPYRIDAE Photuris pennsylvanica (D25416) 1.00/100 Photurinae Cyphonocerus ruficollis (AB604789) Psilocladinae 0.52/99 Drilaster axillaris (AB604790) 0.52/- Stenocladius azumai azumai (AB644225) Ototretinae Photuris pennsylvanica (U31240) Photurinae 1.00/100 1.00/100 Luciola cruciata LcLuc2 (AB490793) 1.00/100 Luciolinae Aquatica lateralis LlLuc2 (AB693934) luciferase Beetle Phrixothrix hirtus head (AF139645) 1.00/100 Phrixothrix vivianii lateral (AF139644) 1.00/100 Phrixothrix hirtus lateral 1.00/100 PHENGODIDAE Brasilocerus sp. lateral (FJ545728)* 1.00/100 (AF139645)* 1.00/88 Taximastinocerus sp. lateral (GU062703)* Rhagophthalmus ohbai (AB255748) 1.00/64 RHAGOPHTHALMIDAE 1.00/100 Pyrophorus plagiophthalmus dGR (AF543413)* 1.00/- Pyrophorus mellifluus dGR (AF545854)* 1.00/98 Pyrophorus plagiophthalmus vOR (AF543373)* 1.00/-Pyrophorus mellifluus vOR (AF545853)* 0.81/- 1.00/66 Pyrophorus angustus dPaLuc (AB479112) ELATERIDAE 1.00/100 0.81/- Pyrearinus termitilluminans (AF116843) Photophorus jansonii (AB767301) 1.00/95 Pyrophorus angustus vPaLuc (AB479113) 0.99/66 Agrypnus binodulus AbLL (AB353886) 1.00/98 Pyrophorus angustus PaLL (AB479114) Luciferase-like 0.67/94 Luciola cruciata LcLL2 (AB196456) Tribolium castaneum TmLL-1 (AB098615) acyl-CoA synthetase 0.67/- 1.00/100 Tribolium castaneum TmLL-2 (AB098616) 0.97/- Luciola cruciata LcLL1 (AB196455) Drosophila melanogaster CG6178 (NM_142964) Oba, Y (2014) Insect Bioluminecence in the Post-Molecular Biology Tribolium castaneum TmLL-3 (AB098617) Era. In: Hoffmann KH (ed.) Insect Molecular Biology and Ecology, Parsley 4-coumarate:CoA ligase 1 CRC
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