Honeybee Cytochrome P450s from version 2 of the Apis mellifera genome

 

Last modified Feb. 22, 2005

 

These sequences were submitted Feb. 11, 2004 by May Berenbaum in collaboration with

Gene Robinson (genome sequencing), Hugh Roberstson (genome annotation) and Reed

Johnson (P450 annotation).

 

On Sept. 2, 2004 Reed Johnson submitted the revised sequences from Honeybee Version

1.2.  These sequence were further edited and revised by D. Nelson on Sept 13-17.

Many partial sequences have been combined and the number of complete genes

increased from 20 to 33.  The number of P450 genes including fragments for ver 1.2

is 55, including 22 partial sequences. 

 

On Feb. 9, 2005 Reed Johnson submitted the revised sequences from Honeybee Version 2.

More sequences are complete and some contamination from cow DNA was noted (CYP2E1).

The gene statistics are 44 complete genes, 3 incomplete but expected to be full length, 2 pseudogenes plus 5-6 small fragments.

 

There are 4 CYP clans in insects.  CYP2, CYP3, CYP4 and mitochondrial

CYP2 is the clan with CYP18 in it.  Sometimes it is called the CYP18 clan.

CYP2 has CYP15, 18, 303, 304, 305, 306, 307, 342, 343

CYP3 has CYP6 and 9 in it and CYP28, 308, 309, 310, 317(a CYP6 subfam)

CYP4 has CYP4, 311, 312, 313, 316, 318

the mito clan has CYP12, 49, 301, 302, 314, 315

 

The honeybee sequences have been sorted into these main CYP clan bins.

48 genes have been named. 1 named gene is a pseudogene.

 

CYP2 clan (8 sequences, 7 complete, 1 incomplete)

 

>CYP18A1 Am2_13.1 (31931-34244), 524 aa, 5 exons

version 1.2 = CYP18A1 Am1.2_13.1b (32196-34509) plus, 524 aa, 5 exons

version 1.1 = Am1.1_13.1a (71904-74217) plus, 524 aa, 5 exons (71904-72236,

72653-72774, 72894-74323, 73404-73668, 73794-74217)

60% to CYP18A1 D. melanogaster

MGGTRIEVLCTFLVFLGVLLVARCLQWLRYVRSLPPGPWGVPVFGYLPFLKGDVHL

RYGELAKKYGPMFSARLGTQLVVVLSDHRTIRDTFRREEFTGRPHTEFINILGGYG(1)

IINTEGAMWKDQRKFLHDKLRGFGMTYMGGGKKIMESRIM(0)

REVKTFLRGLASKRGTPTDVSASLGMSISNVICSIIMGVRFQHGDARFKRFMDLIEEGFKLFGSMAAVNFIPV

MRYLPCLQKVRNKLAENRAEMAGFFQETVDQHRATFDEGTMRDLVDAYLLEIEKAKGEGRATTLFQGKNHD(1)

RQMQQILGDLFSAGMETVKTTLEWAIILMLHHPDAAIAVQEELDQVVGKS

RMPVLEDLPFLPITEATILEVLRRSSVVPLGTTHATTR(2)

DVTLHGYTIPAGSQVVPLLHAVHMDPELWEKPEEFRPSRFLSAEGKVQKPEYFMPFGVGRRMCLGDVLARMEL

FLFFSSLMHTFELRSPQGSSLPSLRGNAGVTVTPDPFDVCLLPRNLDLIEDNDMISTGAILRNIGSH*

 

>Am2_Un.5851 new (1417-191), this is a Bos taurus CYP2E1 low similarity to CYP18 mid region

(1)IIFNNGSTWRDTRRFSLTTLRDLGMGKQGNEQRIQREAHFLLEVLRKTQG(1)QPFDPTFVVGFAPYNVISDILFHKRFDYKDQTSLRLMSLFNENFYLLSSPWIQVKHSPRFCLFCTLLQFDD(1)YLQYLPGSHRKLLKNVSEVKSYALERVKDHQKSLEPSCPRGFLDTMLIEMAKVHS

 

>gi|2511604|emb|AJ001715.1|BTCYP2E Bos taurus mRNA for cytochrome P450 (CYP2E)

          Length = 1603

 

 Score =  313 bits (803), Expect = 2e-84

 Identities = 157/173 (90%), Positives = 160/173 (92%)

 Frame = +3

 

Query: 1   IIFNNGSTWRDTRRFSLTTLRDLGMGKQGNEQRIQREAHFLLEVLRKTQGQPFDPTFVVG 60

           IIFNNGSTWRDTRRFSLTTLRDLGMGKQGNEQRIQREAHFLLEVLRKTQGQPFDPTFVVG

Sbjct: 360 IIFNNGSTWRDTRRFSLTTLRDLGMGKQGNEQRIQREAHFLLEVLRKTQGQPFDPTFVVG 539

 

Query: 61  FAPYNVISDILFHKRFDYKDQTSLRLMSLFNENFYLLSSPWIQVKHSPRFCLFCTLLQFD 120

           FAPYNVISDILFHKRFDYKDQTSLRLMSLFNENFYLLSSPWIQ+ ++           F

Sbjct: 540 FAPYNVISDILFHKRFDYKDQTSLRLMSLFNENFYLLSSPWIQLYNN-----------FP 686

 

Query: 121 DYLQYLPGSHRKLLKNVSEVKSYALERVKDHQKSLEPSCPRGFLDTMLIEMAK 173

           DYLQYLPGSHRKLLKNVSEVKSYALERVKDHQKSLEPSCPRGFLDTMLIEMAK

Sbjct: 687 DYLQYLPGSHRKLLKNVSEVKSYALERVKDHQKSLEPSCPRGFLDTMLIEMAK 845

 

>CYP306A1 Am2_13.1 (38911-36903), 499 aa, 7 exons

version 1.2 = CYP306A1 Am1.2_13.1a (39182-37170) minus, 499 aa, 7 exons

version 1.1 = CYP306A1 Am1.1_13.1b (76876-78884) minus, 499aa, 7 exons (78884-78557,

78493-78345, 78209-78007, 77901-77595, 77538-77383, 77298-77123,

77057-76876)

44% to 306A1 Anopheles gambiae = CYP306A1 in 18clan/2clan

CYP306A1 is the probable ortholog of CYP306A1 in diptera (flies and mosquitos)

the % identity is below the usual cutoff for subfamily membership,

but it makes sense to name orthologs with the same name.

MILDHYIAIFVLPFLLLLYVVRKNRKARRLPPGPWQLPLLGYLPWIDAEKPHETLTRLS

RVYGPVCGFRMGSVYTVLLSDPQLIRQSFAKDSITNRAPLYLTHGIMKGYG(1)

IICAEGEQWKDQRKFISNCLRNFGMVKHEGAKRDKMEERISDAVNECVS(0)

VLRDRGANGPIDPLDTLHHCLGNLVNSIVFGKTYEEEDRIWKWLRHLQEEGVKQIGVAGPLNFLPFLR(2)

FLPQYGRVIRSIVDGKDKTHEIYRQILDEHRARVDSGNGCKIDSFLAAFDEQ

MRKKDGAESGYFTEPQLYHLLADLFGAGTDTTLTTLRWFLLFMAAHPMEQ(0)

EKIQSEMDLCLREGEQPTLNDRIVMPRLEAAIAEVQRIRSVTPLGIPHGTSE(0)

DVEIGGYDIPCGAMIVPMQWAIHTDPAYWRDPLEFRPDRFLSEDGTFFKPESFLPFQNG(1)

KRVCVGEELARMILFLFAGRILRAFSVRVPAGEIADLEGECGITLVPKPHRLAFVGRDR*

 

>Am2_Un.5840 new (1363-466), 74 aa, 2 exons 41% to 306A1 this is a CYP2E seq from Bos taurus

(0)GTVVIPTLDSVLHDRQEFPEPEKFKPEHFLNENGKFKYSDHFKAFSAG(1)KRVCVGEGLARMELFLLLAAILQHFT*

 

>gi|2511604|emb|AJ001715.1|BTCYP2E Bos taurus mRNA for cytochrome P450 (CYP2E)

          Length = 1603

 

 Score =  123 bits (308), Expect = 3e-27

 Identities = 62/73 (84%), Positives = 62/73 (84%)

 Frame = +3

 

Query: 1    GTVVIPTLDSVLHDRQXXXXXXXXXXXHFLNENGKFKYSDHFKAFSAGKRVCVGEGLARM 60

            GTVVIPTLDSVLHDRQ           HFLNENGKFKYSDHFKAFSAGKRVCVGEGLARM

Sbjct: 1176 GTVVIPTLDSVLHDRQEFPEPEKFKPEHFLNENGKFKYSDHFKAFSAGKRVCVGEGLARM 1355

 

Query: 61   ELFLLLAAILQHF 73

            ELFLLLAAILQHF

Sbjct: 1356 ELFLLLAAILQHF 1394

 

>CYP307B1 Am2_14.4 (871542-875286), 507 aa, 3 exons

version 1.2 = CYP307B1 Am1.2_14.10 (757965-761727) plus, 507 aa, 3 exons

version 1.1 = Am1.1_14.9 (18918-22680) minus, 507 aa, 3 exons (22680-22278,

20154-19363, 19246-18921)

CYP307B1  55% to 307B1 Anopheles, probably the ortholog of 307B1

MIPLTATTCFLIAITFLALALILLDHLRSKKTTKSVVPGDDDQHALPEPPGPKPWPILGSLHILGRYDVPYKAFADLVRDFDCQVIKLRMGSVPCVVVNGLENIKEVLTVKGHHFDSRPNFARYHLLFGGNKENS(1)

LAFCNWSDVQKARREMLRAHTFPRAFSTRFNELNGIIGDEMEFMVNHLDSLSGTSVHAKPLILHCCANIFITYLCSKNFHLEHDGFRNMVENFDKVFFEVNQGYAADFLPFLMPLHHRNMARMAHWSHEIRRFVIKNIIADRVNSWNDVVPEKDYVDCLINHVKSGTEPQMSWNTALFVMEDIIGGHTAIGNLLVKVLGFLATRPEIQRLAQDEIDALGLAGNFVGLENRRSLPYVEAIILETIRIIASPIVPHVANQDSSIAG(1)

FRIKKDTFIFLNNYDLNMSTDLWTSPEEFMPDRFVQNGRLLKPEHFLPFGGGRRSCMGYKLVQYVSFAILASILKNFTITPVQKEDYTIPIGNLALPEMTYKFRFERR*

 

>CYP303A1 Am2_Un.6309 (6408-4574), 375 aa, 8 exons

version 1.2 = CYP303A1 Am1.2_Un.1361 incomplete on N-terminus (5389-7223) minus, 378 aa, 8 exons

AADG03013520.1 (WGS)

version 1.1 = CYP303A1 Am1.1_Un.2253 incomplete on N-terminus (55-1889) plus, 378 aa, 8 exons

43% to 303A1 D.mel., 44% to 303A1 Anoph. CYP303A1 in 2 clan/18 clan

may be the ortholog of 303A1, first exon below is 75% to 304A1

(1)LLLVDGNLWNEQRRFVLKHLRDFGFGRQN (1)

LYMNANEYTGNNVTQSQLGTIISMHNIFGITVLNSLWKMLAGKR (2)

YNIDDKELIYFQRILSITLNEIDMLGAPFSHFPLLRFIAPEISGYKSFVKIHEELWKFFK (0)

DEVNNHKNTFNSDSPGNLIDIYLTILNSENYGKTFSD (1)

VSEPQLVAICVDLFMAGSETTSKVLGFCFLYLVLFPHVQKKAHEEIDRVIGRNKLPTAEDKAK (2)

MTYMNAIVLESLRMFAGRSLNLPHRVQRDTKISDYKIPK (0)

NTIIITNFNGILMDESWGDPENFRPERFIDGSGNIVTPSRFLPFSAG (1)

KHRCMGENLAKTNIFIIATTLLQAFTFSEIPGEKPTIEHFIDGTTISPKPYRVNVSLRI*

 

>CYP305D1 Am2_7.2 frameshift repaired in 7th exon  (72431-68935), 490 aa, 8 exons

version 1.2 = CYP305D1 Am1.2GroupUn.127b (70044-73538) minus, 490 aa, 8 exons (first exon is a guess)

version 1.1 Am1.1_Un.7452 incomplete on both ends (700-954) minus, 85 aa, exon 2

version 1.1 Am1.1_Un.6110 incomplete on both ends (1-622) minus, 126 aa, exons 6,7

Two fragments from v.1.1 have been combined in this sequence.  The first exon is a best guess. BI513047.1 (EST) AADG03008281.1 (WGS) 39% to 305A2, 39% to 305B1 Bombyx

39% to 305B1 silkworm

MNKNFVKIFNLLLYLFIDVFNNLFFIECDG(1)

PFSWPFIGNQILLKRLSRKFGGQHKAFMELSKRYNSDIITVNISYEKIIVVSGSK

FCDMILQNEEFQGRPWNEFIKVRNMGKKQG(1)

ITMNDGTEWKELRNWMMRTMKIFGFGKSEMIEMIQHQLVIFSENLNKNKLHQLKLLFVPAVINVLWNFITGELVAFNQQQK(2)

LEHFLDLLDRRSRCFDITGGLLAAFPWIRYIAPEISGYNIMCMLNKELKDFLM(0)

KTINDHKEKYIEGKEADLIDMFIQEMRKNEKSSIFTD(1)

EQLMMILIDLFLAGFTTTSTTLDFLFLIVTLFPDVQRKVQKEIDSVIPYDRLPNMEDKAK(2)

LPYVEAVISETYRLWPVFPIIGPRRVLCDTNIDKYVIPKDTTILFNTYSINKDPTL

YPDPDKFMPERFIKNGVFEPDEYSLQFGKG(1)

KRRCPGDILAKATIFILFVGIMQKYTLLPVPGKGPHSIKINSGITLTPQPYNVLVEKR*

 

>CYP15A1 Am2_7.2  (65177-67812), 500 aa, 8 exons

W->L, changed splice donor on penultimate intron and both splice sites on final intron

version 1.2 = CYP15A1 Am1.2_Un.127a (66491-68918) plus, 504 aa, 8 exons XM_392687.1 partial

gnl|Amellifera1|165771382 BCM Apis mellifera 12/11/2003

Am1.1_Un.897 incomplete, missing exon 5 (51606-53773) plus, 357 aa, 5 exons

Am1.1_Un.10970 incomplete on N-terminus (112-677) plus, 121 aa, 2 exons

Two fragments from v.1.1 have been combined.  18clan/2clan

47% to CYP15A1 Diploptera punctata (probable ortholog)

39% to CYP15B1 Anopheles gambiae 36% to 303A1, 35% to 305A3, 31% to 304B1

MLYVVISLLLALYCIFCIYDCVKPHNFPP (1)

GPKWLPLIGCFLTFRRLKLKHKYTYVAFQELSKTYGPILGLKLGSQKLVVI

STHDLVKKVLLQDEFNGRPDGFFFRVRAFGKRK (1)

GILFTEGSMWSQCRRFTMRHLRSFGLGQSTMEKYLTVEAENLVNYLRRVST

KGPVPMHTAFDIAVLNSLWCMFAGHRFDYENEKLAEILEIVHDSFR (2)

LMDTMGGIISQMPFLRFIIPELSGYNNLMEILRKLWNFLDEEINNHEKHLSGNQPQDLIEAFLLEISSRNGVQNDSIFDR (1)

ENLLILCLDLFLAGSKTTTDTLSTSILFLSLHSEWIKILQEELDNVVGRSRSPTLEDYSSLPIMESFLAE (0)

IQRFLILAPLGVPHKTTKDVILNGYNIPK (0 GC boundary?)

DTTVLLDFHSAHNDPAYWDHPEEFRPQRFLDANGRFCQNNANIPFGL (1)

GKRRCPGEMLARTSLFLYFAYVIHYFDIEISPEHGKPDLNGHDGFTISPKSYYLKITARSDVTNCSTI*

 

Note: I still argue for a GC boundary at YNIPK, because this is a conserved sequence and it should not be deleted to make a conventional GT boundary at TTKD. The same is also true at the heme signature intron.  A GC boundary preserves conserved motifs at this site.

 

>Am2_Un.4815  new (4673-4457), 72 aa, 1 exon, similar to 15A1 this is Bos taurus CYP2E1

(2)MGLPWRQRGSPHGLLSLQLAERYGPVFTLYLGSQRAVVVHGYKPVKEVLLDYKNEFSGRGENPGFQMHKNNG(0)

 

>CYP2E1 TC189658 = AJ001715 79% to human 2E1 Length = 1900

        Length = 496

 

 Score = 291 (102.4 bits), Expect = 7.1e-29, P = 7.1e-29

 Identities = 54/55 (98%), Positives = 55/55 (100%)

 

Query:    18 QLAERYGPVFTLYLGSQRAVVVHGYKPVKEVLLDYKNEFSGRGENPGFQMHKNNG 72

             +LAERYGPVFTLYLGSQRAVVVHGYKPVKEVLLDYKNEFSGRGENPGFQMHKNNG

Sbjct:    59 RLAERYGPVFTLYLGSQRAVVVHGYKPVKEVLLDYKNEFSGRGENPGFQMHKNNG 113

 

>CYP342A1 Am2_8.3 (44357-46424), 505 aa, 7 exons

version 1.2 = CYP342A1 Am1.2_8.3 incomplete on N-terminus (1269-3097) plus, 505 aa, 7 exons

version 1.1 Am1.1_Un.8493 incomplete on both ends (509-1028) minus, 141 aa, 2 exons

36% to 304A1 36% to 304B1 18clan/2clan

MISFLFIIFLLLIIYKIYNSVIHVSSNTPPC(1)

LPRLPIIGSYWHLLWHDYEYPYNGIIHYVNKLQSKIVTCYFGSHKTIIANDYKSIKEVL

TKQEFNGRPINVDIVLQRAFGKSLG(1)

IFFTEGTLWHEQRRFALRHMRDFGFGRRHEIFETNVMEEIAILVDMLKEGPINDEEK(0)

KFLKNGYACFPDILYPYVANVILNIMFGERFDRSQYHKLIYFCESSMMFQKSLDTSG

GAIFQFWFLKYFGNIFGYTNAIKATYQMINFIE(0)

EYIDNKKDLDDYDKGLIGRYLKILKEKNNITSTFSQKQLIMTLVDFMFPATSALPSA

LVHAIKLVMHHPRVVNNIQEEIDRVVGTGRLVTWSDRKN(2)

LPYIEATIRESLRYETLTPLSVFHKTLKKTTLCDYDIPKDTLVVTNLVALNTDPDLW

GDPENFRPERFLDENNELRKDFTFPFGFG(1)

HRVCPGETYSRYNMFEVFAVLMQNFNFSFVEGEPTGLDDKESGLIVTPKKTWIQVKARNMK*

 

>CYP343A1 3241+6423 Am2_Un.573, R->K (725-7080), 496 aa, 8 exons  combined Am1.2_Un.3241 and Am1.2_6423 to complete

version 1.2 = Am1.2_Un.3241 (94-459) minus, 96 aa, 2 exons at N-terminus up to C-helix

version 1.1 = Am1.1_Un.7901 incomplete on C-terminus (1-459) minus, 126 aa, 2 exons 40% to 15B1

version 1.2 = Am1.2_Un.6423 incomplete on N-terminus (67-247) minus, 254 aa, 4 exons

AADG03019874.1 (WGS)

Am1.2_Un.1305 incomplete on N-terminus (2043-2647) plus, 146 aa, 2 exons

Am1.1_Un.960 incomplete on both ends (8311-11562) minus, 147 aa, 2 exons

One fragment in v.1.1 was divided into two in v.1.2, but they are probably from

one gene. 34% to 305a4, 36% to CYP15A1, new family in the CYP2 clan

MWFVILCFVIVLIKILFDYSRPINFPPG(1)

PRGLPFIGNILDIIKLINETKYYSDTWCRLAEKYGSVVGLRLGLDQPLIIVSGKSAVTEMLNR

SEFDGRPSGFLYKYRCGGMQQGILFTDTDVWHSQRR(2)

FALKTLKQFGFGKNSMEHILQHDAIALTNIIIELTKDGTVKNIRSIISAAVLSNLWLLIDGTK(2)

FDIGMENSNLKEAINIVQDIVKSSNVSGGIINQFPFLRHLFPNLTGFSAFVERQKRINNFFM(0)

EVIAKHKWKKINEEGTNFIDVYLQEIQKKNSSHSFFNE(1)

NQLLYIIKDLFSAGVDTTNSTIGFIIAFLVVHQDVQSKVYDEISRVIDKDIYPSLSDKDR(2)

LPYLKAVIAEVSRLANIGPTSIPHRAVKDSTFLGFEIKKNYTLLANFKSIHMDKEHWGDPEIF

RPERFINEKGDFINDSWLMPFGLG(1)

RRKCLGETLAKNTVFLFVACMLQRLHFMLPSNHPPPCLQGIDGFVIAPPMMDIIAVQRF*

 

note: intact seq 36% to 15A1, 35% to 15B1, 34% to 305A4

 

CYP3 clan (31 seqs, 28 complete and 3 incomplete, 2 of the incomplete are pseudogenes, 5-6 fragments)

 

>CYP6AQ1 Am2_12.16   (1067523-1070348), 514 aa, 5 exons

version 1.2 = CYP6AQ1 Am1.2_12.14 (417204-420026) plus, 514 aa, 5 exons

version 1.1 = CYP6AQ1 Am1.1_12.14 (315628-318450) plus, 514 aa, 5 exons (315628-316108,

316238-316604, 316677-316861, 316989-317277, 318272-318450)

45% to 6K1, 42% to 6g2m 43% to 6G1ps new subfamily in CYP6

cyan = missing seq. from EST BE844578

yellow = EST BE844462, underlined seq = EST BE844394, green = EST BE844353

magenta = EST BE844352, gray = EST BE844331 all ESTs from antennae

 

MNLLTPYWSLDILIVSSSLMIAVYLYASWKLKYWSRRGIMQITPSPLFGNFKKCILFQKSVSEIIRELYGQNEGLPFMGFY

IFYKPFFLVRDIELVKHILVKDFNTFANKHTSADSKNDRIGYSNLFIIKNPAWKYLRGKLTSVFTSGKLKKMFDLMLIIG (1)

KNLEKHLELLNLDG

NGKEVELKDLCANFTTDLIGTTAFGVNLNSLKDPNSDFRENGRLVFDYNLKRAFEFFSIFFFPNLS

KYVSIKFFGKATDYFRNSFWSVINQRIESNVKRNDLIDCLIELREKHKNDESFEGFR (1)

FDGDDLVSQAAIFFTGGFETSSTTISFTLYELALNKDIQKTVRTEIHEALAQTDGKITYDM (0)

ITNLPYLDMVVSETLRKYPPLGFLDRVALHDYKIPNSDVTIDKDTPVIIPMIAFHYD

PKYFPNPEKYDPLRFSEEVKKTRPSYVYMPFGEGPHICIG (1)

MRLGLLQSKLGIIEILKDYEVSPCEKTKIPMVLDPKGLTTTALGGLYLNIRKITIAAG*

 

>CYP6AR1 Am2_5.8 (434944-432195), 502 aa, 5 exons

version 1.2 = CYP6AR1 Am1.2_5.5 (430078-432827) minus, 502 aa, 5 exons

version 1.1 = CYP6AR1 Am1.1_Un.19 (44801-47550) plus, 502 aa, 5 exons

50% to AmGroupUn.5496, 47% to AmGroupUn.792b, 38% to 6a13ps all best hits to 6as

MSWLMIETVGLIATVFFLLYYYSMSKLDYWRKRGVKGPKPLPFLGNFKDVLLAKESTMDCFERA

YKEFKDEPMVGMYGSHEPLLILRDLDLIKDVLIKDFNKFAQRTQGAIRE(0)

VEPLSEQLFRLDAERWRPLRLKLSSFFSSGKLKEMFHLFVECSDNFEKYLEKMVEKGGLVECRDAAAKFSTDVIGACAFSIHTNALTDENSQFRKMGKQALATNLQQFLNDRLREYPFLFKIFGRFFVDHEVTNFFANSIKDAMDY

RIQNNVHLRDVIDILADIRENPTKCGLKE(1)

ADNLFLTSQAVLFFLAGFENASLTISNALYELAWKPEIQEKARAEIVNVLQKYDGKITYDGLEEMKYLEACIFE(1)

TLRMYPVLQWLSREAMETYTFTGTKVTIPKGQQVFLPIYAIQRDPDI

YPNPDNFDPERFTDDKIKTRHSMTHLPFGDGPRHCSG(1)

IRLAKKQLKVGLVTVLSKFKVEVCEKTRKIYQKDKKPLFLLQPVDGIHLKISKVSV*

 

>CYP6AS1 Am2_Un.5363 (3492-1105), 498 aa, 5 exons

version 1.2 = CYP6AS1 Am1.2_Un.6491 (264-2651) plus, 498 aa, 5 exons

version 1.1 = CYP6AS1 Am1.1_Un.5496 (264-2651) plus, 498 aa, 5 exons

44% to 6A14, 64% to AmGroupUn.792b

MDYFQILCAISIVILTIYYYYSSKYTFWKKRGISGPKPIIFFGNFVDSIIQKRSTSEAVKK

WYDDYKHESVFGIFGGTTPLLVINDLDMIKDVLIRDFSLFVDRGFHIFPK(0)

IEPLSEHLFLLEAERWRPMRMKLSPIFTSGKLKEMFFLIMESAGNLEKYLDEVIKKDEMVECRELAAKFMTDVIGSCAFGINTNSLLEEDSEFRRMGKKISTPNLKVMLGNICKEFFPPLYEIVGSIFTLKDVNEFFIN

LVSDTMKYRKDNNIIRSDFINMLMQLKEHPEKMENIE(1)

LTNTLLTAQAVVFFVAGFETSSSTMAFSLYELAQNQEIQDKLREEIRNMHEKNKGVLTYTDVKEMKYLDKVFKE(1)

TLRKYPILPMLFRQAMENYTFKDTKITIPKGMKLWVPVHGIHHDPNI

YPEPEVFDPERFEDDAFASRHPMSYLPFGDGPRNCIG(1)

ARFAHYQSKVGLITILRHHKVNVCEKTTIPFKADERSFLLTLKGGVHLKITKI*

 

>CYP6AS2 Am2_Un.476 (25830-23485), 498 aa, 5 exons

version 1.2 = CYP6AS2 Am1.2_Un.601a (2171-4675) plus, 498 aa, 5 exons

version 1.1 = CYP6AS2 Am1.1_Un.792a incomplete on C-terminus of exon 2, missing 3rd exon

(8375-10702) minus, 356 aa,  EST BE844607 from antennae

86% to AmGroupUn.5496

MDYFQILCAISIVILTIYYYYSLKYAFWKDRGISGPKPIIFFGNFGNSIIKKKSLSETVK

KWYDDYKHESVFGIFEGTIPVLVINDLDMIKDILIRDFSLFVDRGFHIFPK(0)

IEPLTQHLFLLEAERWRPMRMKLSPIFTSGKLKEMFSLIVESAGNLEKYLDEVIKKNEMVECRDLAAK

FTTDVIGSCAFGINTNSLLEEDSEFRRMGKKIFSPSLKLMIGNTCKVFFPSLYEVIGNIFTMKDVDEF

FINLVSDTMKYRKDNDIVRSDFINMLMQLKEHPEKMDNIE(1)

LTDTLLTAQAVVFFIAGFETSSSTIAFGLYELAQNQEIQDKLREEIRKMHEKNKGILTYTDIKEMKYLDKVFKE(1)

TLRKYPILSTLSRKAMENYTFKGTKITIPKGTKVWVPVYGIQHDPNI

YPKPEVFDPERFEDDAFASRHPMSYLPFGDGPRNCIG(1)

ARFAHYQSKVGLITILRNHKVNVCEKTTIPFKADERSFLLALKGGVHLKITKI*

 

>Am2_Un.4631 new (605-937), 111 aa, 1 exon

only one aa diff to 6AS2 probably the same seq.

MDYFQILCAISIVILTIYYYYSLKYAFWKDRGISGPKPIIFFGNLGNSIIKKKSLSETVKKWYDDYKHESVFGIFEGTIPVLVINDLDMIKDILIRDFSLFVDRGFHIFPK(0)

 

>gi|15354903|gb|BI504529.1|   BB170023B10C08.5 Bee Brain Normalized/Subtracted Library, BB17 Apis

           mellifera cDNA clone BB170023B10C08 5', mRNA sequence.

          Length = 657

 

 Score =  190 bits (483), Expect = 1e-48

 Identities = 88/114 (77%), Positives = 97/114 (85%), Gaps = 7/114 (6%)

 Frame = +3

 

Query: 18  YYYYSLKYAFWKDRGISGPKPIIFFGNLGNSIIKKKSLSETVKKWYDDYKHESVFGIFEG 77

           YYYYS KYAFWKDRGISGPKPI+FFGN GNSI+KK+S+SETVKKWYDDYKHESVFGI+EG

Sbjct: 3   YYYYSSKYAFWKDRGISGPKPIVFFGNFGNSIVKKRSISETVKKWYDDYKHESVFGIYEG 182

 

Query: 78  TIPVLVINDLDMIKDILIRDFSLFVDRGFHIFPK-------LLLVDGNLWNEQR 124

           TIPVLVINDLDMIKD+LIRDFS+FVDRGFH FPK       L L++   W   R

Sbjct: 183 TIPVLVINDLDMIKDVLIRDFSIFVDRGFHTFPKIEPLTQHLFLLEAERWRPMR 344

 

an EST  95% to 6AS2 above, exact match to Un.601a from ver 1.2

There might be alternative splicing of the first exon.

YYYYSSKYAFWKDRGISGPKPIVFFGNFGNSIVKKRSISETVKKWYDDYKHESVFGIYEG

TIPVLVINDLDMIKDVLIRDFSIFVDRGFHTFPKIEPLTQHLFLLEAERWRPMRMKLSPI

FTSGKLKEMFSLIVESAGNLEKYLDEVIKKNEMVECRDLAAKFTTDVIGSCAFGINTNSL

LEEDSEFRrMGKKIFSPSLKLMIGNTCKVFFPSLYEVI

 

>CYP6AS13 CYP6ASf1 Am2_Un.5370 (9442-11518), 497 aa, 5 exons,  completion on C-terminus stop codon changed to A (63% to 6AS2)

version 1.2 = CYP6AS fragment Am1.2_Un.1326b incomplete on C-terminus (4-243) minus, 80 aa, 1 exon

1 aa diff to Am1.2_Un.5413

version 1.2 Am1.2_Un.5413 stop codon in 3rd exon, incomplete on both ends, (1-1334) plus, 315 aa, 3 exons 60% to 6AS2, 71% to Am1.2_Un.6932 in one overlapping exon 3

MDFFQIFCAICIMLLAIYYYYTSIYNYWKVRGIPGPEPTIIIGNFMEVFLKKISINDKLRFL

YNKYKNEPMFGIFEGSSPILVLNDLDLIKDVLIKDFSIFSNRGFRIFPK(0)

AEPLGEHLFALETERWRPMRAKLSPIFTSGKLKEMFPLIIECSKNMEPYLDKIAERGKYIECRDLAAKFTTDVIGSCAFGIDMNSISDKDSEFRIIGRKLFTPTFKTIVRDVCRQFLPGLYDVIGHKLQIEEVNEFLT

NLIKDTINYRKENKIVRPDFVNTLIELKDHPEKLETIK(1)

LTDSMIASQAFVFFVAGFETSSSTISHALYELAQNQEIQDKLREEIREVYEKHGELTYDVIKNMKYLDKVLKE(1)

TLRKYPIMAMLTREAQENYTFKGTKVTIEKGIKVWILPYGIQNDPDIFPNPDI

FDPERFDEEAVAARHPMSYLPFGDGPRNCIG(1)

ARFAQFQSKIGIITIVRNHKIDVCEQTKIPYESDPFQFLLALKGGINLKISKI*

 

>CYP6AS2P1 CYP6ASf2 Am2_Un.5353 (2878-1897), 209 aa, 3 exons,  4 aa change at C-end of last exon due to frameshift, frameshift in middle of last exon

version 1.2 = CYP6AS fragment Am1.2_Un.6932 incomplete on N-terminus (41-1021) minus, 209 aa, 3 exons, 92% to last 3 exons of CYP6AS2

(1)LTDILLTAQAVVFFVAGFETSSSTMAFSLYELAQNQEIQDKLREEIRKVHEKNKGVLTYTDIKEMKYLDKVFKE(1)

TLRKYPILSTLSRKVMENYTFKGTKITIPKGTKIWVHGIQHDPNIYPEPEVFDPERFEDDAFASRHPMSYLPFGDGPRNCIG(1)

LVLPHYQSKVGLITILRNHKXVNVCEKTTIPFKADERSFLLVPKGGIHLKIIKI*

 

6AS2 (top) compared to 6ASf2

Query: 1   LTDSMIASQAFVFFVAGFETSSSTISHALYELAQNQEIQDKLREEIREVYEKH-GELTYD 59

           LTD ++ +QA VFFVAGFETSSST++ +LYELAQNQEIQDKLREEIR+V+EK+ G LTY

Sbjct: 1   LTDILLTAQAVVFFVAGFETSSSTMAFSLYELAQNQEIQDKLREEIRKVHEKNKGVLTYT 60

 

Query: 60  VIKNMKYLDKVLKETLRKYPIMAMLTREAQENYTFKGTKVTIEKGIKVWILPYGIQNDPD 119

            IK MKYLDKV KETLRKYPI++ L+R+  ENYTFKGTK+TI KG K+W+  +GIQ+DP+

Sbjct: 61  DIKEMKYLDKVFKETLRKYPILSTLSRKVMENYTFKGTKITIPKGTKIWV--HGIQHDPN 118

 

Query: 120 IFPNPDIFDPERFDEEAVAARHPMSYLPFGDGPRNCIGARFAQFQSKIGIITIVRNHK-I 178

           I+P P++FDPERF+++A A+RHPMSYLPFGDGPRNCIG     +QSK+G+ITI+RNHK +

Sbjct: 119 IYPEPEVFDPERFEDDAFASRHPMSYLPFGDGPRNCIGLVLPHYQSKVGLITILRNHKXV 178

 

Query: 179 DVCEQTKIPYESDPFQFLLALKGGINLKISKI 210

           +VCE+T IP+++D   FLL  KGGI+LKI KI

Sbjct: 179 NVCEKTTIPFKADERSFLLVPKGGIHLKIIKI 210

 

the bottom seq has some defects.  The last frameshift X should not be counted as an extra amino acid since it adds to the length and this does not match 6AS2.

 

The heme region is out of phase

 

     1: K  C  I  F  D  A  V  F  L  *  I  L  I  L  I  F  V  I  I  F 

     2:  N  A  F  S  M  Q  F  F  Y  K  F  *  F  *  F  S  *  L  F  F

     3:   M  H  F  R  C  S  F  F  I  N  F  N  F  N