Les phéromones de la Drosophil hile : évoluti tion et rôle dans la spéciation e i Wicker Th omas

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1 Les phéromones de la Drosophile : évolution et rôle dans la spéciation Claude Wicker-Thomas legs, UPR 9034, Gif sur Yvette

2 Drosophila courtship pheromones pheromones pheromones pheromones

3 Pheromones are synthesized in the oenocytes oenocytes Abdomen (lateral view) desat1 (a pheromone biosynthesis gene) expression Wicker-Thomas, Chertemps In Insect hydrocarbons : biology, chemistry and chemical ecology, in press.

4 Main cuticular compounds D. simulans and D. melanogaster 7T Pheromonal compounds 7P 7-T 7 23 D. melanogaster 7,11HD 7,11ND 7-P 7 25 C23 C25 C27 7H C29 Time (min.) 7,11-HD 7,11-ND

populations Mate less rapidly Sexual isolation Jallon and Péchiné

5 Geographical pheromone polymorphism in D. melanogaster females 7,11-HD/5,9-HD 5,9-HD flies Only a few populations Mate less rapidly Sexual isolation Jallon and Péchiné 1989 CRAS; Ferveur and Jallon 1996 Genet Res; Rouault, Marican, Wicker-Thomas, Jallon 2004 Genetica

6 Hydrocarbon biosynthesis (D. melanogaster) elongation decarboxylation C14 C16 C2n C2n-1 Saturated hydrocarbons No pheromonal role ω5 ω7 Δ9 desaturation No pheromonal role elongation decarboxylation C14 :1 C16 :1 C2n:1 C2n-1:1 Desaturation and elongation decarboxylation C2n:2 C2n-1:2 Main hydrocarbons in males 7-T Main hydrocarbons in females 7 7-P 7 7,11-HD 5,9-HD 7 11 Cosmopolitan strains 5 9 African strains Wicker-Thomas, Henriet, Dallerac 1997 IBMB Dallerac, Labeur, Jallon, Knipple, Roelofs, Wicker-Thomas 2000 PNAS

7 Properties of desat locus desat1 is expressed in all strains (males and females) desat2 is only expressed in «5,9-HD» morph females Desat1 MPPNAQAGAQ SISDSLIAAA SAAADAGQSP TKLQEDSTGV LFECDVETTD GGLVKDITVM Desat MAPYSRIYH QDKSSRETGV LFEDDAQTVD SDLTTDRFQL Desat1 KKAEKRLLKL VWRNIIAFGY LHLAALYGAY LMVTSAKWQT CILAYFLYVI SGLGITAGAH Desat2 KRAEKRRLPL VWRNIILFAL VHLAALYGLH SIFTRAKLAT TLFAAGLYII GMLGVTAGAH NH 2 COOH Desat1 RLWAHRSYKA KWPLRVILVI FNTIAFQDAA YHWARDHRVH HKYSETDADP HNATRGFFFS Desat2 RLWAHRTYKA KWPLRLLLVI FNTIAFQDAV YHWARDHRVH HKYSETDADP HNATRGFFFS CYTOSOL Desat1 HVGWLLCKKH PEVKAKGKGV DLSDLRADPI LMFQKKYYMI LMPIACFIIP TVVPMYAWGE Desat2 HVGWLLCKKH PDIKEKGRGL DLSDLRADPI LMFQRKHYYI LMPLACFVLP TVIPMVYWNE Desat1 SFMNAWFVAT MFRWCFILNV TWLVNSAAHK FGGRPYDKFI NPSENISVAI LAFGEGWHNY Desat2 TLASSWFVAT MFRWCFQLNM TWLVNSAAHK FGNRPYDKTM NPTQNAFVSA FTFGEGWHNY RETICULUM MEMBRANE Desat1 HHVFPWDYKT AEFGKYSLNF TTAFIDFFAK IGWAYDLKTV STDIIKKRVK RTGDGTHATW Desat2 HHAFPWDYKT AEWGCYSLNI TTAFIDLFAK IGWAYDLKTV APDVIQRRVL RTGDGSHELW ER LUMEN 361 Desat1 GWGDVDQPKE EIEDAVITHK KSE Desat2 GWGDKDLTAE DARNVLLVDK SR Membrane protein

ole1 yeast 2 days after transformation at 29 C Heterologous expression

C16:0 C18:1 Desat1 Control ω7 Δ9 C16:1 CH3-(CH2) 5 -CH=CH-(CH2) 7

1 CH3-(CH2) 3 -CH=CH-(CH2) 7 -COOH C14:0 C18:0 Desat2 C14:11 C16:0

8 ole1 yeast 2 days after transformation at 29 C Heterologous expression of desat1 and desat2 desat1 desat2 Fatty acid analysis Desat1 C16:1 C16:0 C18:1 Desat1 Control ω7 Δ9 C16:1 CH3-(CH2) 5 -CH=CH-(CH2) 7 -COOH C18:1 CH3-(CH2) 7 -CH=CH-(CH2) 7 -COOH ω9 Δ9 Desat2 ω5 Δ9 C14:1 1 CH3-(CH2) 3 -CH=CH-(CH2) 7 -COOH C14:0 C18:0 Desat2 C14:11 C16:0 C14:0 C18:0 Dallerac, Labeur, Jallon, Knipple, Roelofs, Wicker-Thomas 2000 PNAS

9 Polymorphism of 43 lines of D. melanogaster Morph Origin Number Presence of Coding region of strains 16nt sequence High 5,9 HD African/Caribbean 15 Yes High 5,9 HD Cosmopolitan 1 Yes Low 5,9 HD Cosmopolitan 18 No Low 5,9 HD African/Caribbean i 5 No Low 5,9 HD African/Caribbean 1 No stop codon Intermediate Cosmopolitan 2 Yes/ No Intermediate African/Caribbean 1 Yes/ No CCAAT TAAAAATA ATTCGGTCGGGCCAAGAA Takahashi, Tsaur, Coyne, Wu 2001 PNAS

10 Promoter prediction for Canton-S desat2 CAAT TATA Position of the 16 nt deletion gif ATG Promoter prediction for Tai desat2 TATA CAAT Position of the 16 nt sequence gif ATG

11 Evolution of 7,11-HD and 5,9-HD morphs Enoyl-CoA Hydratase CCAAT TAAAAATA ATTCGGTCGGGCCAAGAA STOP poly A signal putative TATA box putative 16 nt sequence present in Tai strain initiation absent in Canton-S site exon 1 exon 2 exon 3 exon 4 TAAATATATT STOP poly A signal Ancestral African female Cosmopolitan female genome desat2 desat1 desat2 desat1 Enzymes Desaturase2 Desaturase1 Desaturase1 Hydrocarbons quantities per fly 5,9 HD 7,11 HD 7,11 ND 7,11 HD 7,11 ND 600 ng 100 ng 200 ng 400 ng 200 ng genetic advantage?

12 Desat2 transcription D. melanogaster «7,11-HD» Canton S not expressed (Dallerac et al. 2000;Greenberg et al., 2006) Some strains expressed at a low level (Michalak et al., 2007) D. melanogaster «5,9-HD» Tai expressed only in females (Dallerac et al 2000) Other strains expressed in females ++ expressed in males (from 0 to ++) (Greenberg et al 2006) D. simulans : very little 5-HC expressed in males

13 Questions What does the promoter look like in the Drosophila strains or species which express desat2? (Is the 16nt deletion absent?) If desat2 is expressed in 7-HC flies, where is it expressed? In 5,9-HD strains of D. melanogaster, where is desat2 expressed?

14 Reproductive isolation in Lepidoptera Ostrinia nubilalis (European corn borer) Z11/E11-14:O-Acetate (97:3) Δ11 desaturase Ma et Roelofs (2002) Ostrinia furnacalis (Asian corn borer) Z12/E12-14:O-Acetate (97:3) Δ14 desaturase Klun et al. (1980)

15 Reproductive isolation in Lepidoptera Ostrinia nubilalis (European corn borer) Z11/E11-14:O-Acetate (97:3) Δ11 desaturase 3 mrna present: Δ9,, Δ11 et Δ14 desaturases Ma et Roelofs (2002) Ostrinia furnacalis (Asian corn borer) Z12/E12-14:O-Acetate (97:3) Δ14 desaturase 3 mrna present: Δ9,, Δ11 et Δ14 desaturases Roelofs et Rooney (2003) The 3 enzymes have a desaturase activity in vitro Only one is used for pheromone biosynthesis: Δ11 desaturase in O.nubilalis Δ14 desaturase in Of O.furnacolis

16 Questions What does the promoter look like in the Drosophila strains or species which express desat2? (Is the 16nt deletion absent?) If desat2 is expressed in 7-HC flies, where is it expressed? In 5,9-HD strains of D. melanogaster, where is desat2 expressed?

17 Pheromone biosynthesis in Drosophila Fatty Acids Hydrocarbons Steps common to males and females elongation decarboxylation C16 C2n C2n-1 ω7 Desat1 C16 :1 C2n:1 C2n-1:1 D. simulans D. melanogaster males 7-T 7-P DesatF EloF (female specific) C2n:2 C2n-1:2 Wicker-Thomas, Henriet, Dallerac, 1997, IBMB Dallerac, Labeur, Jallon, Knipple, Roelofs, Wicker-Thomas, 2000, PNAS Chertemps, Duportets, Labeur, Ueda, Takahashi, Saigo, Wicker- Thomas, 2007, PNAS D. melanogaster females 7,11-HD Steps specific to D. melanogaster and D. sechellia females

18 QTL analysis of hydrocarbons differing between D. simulans and D. sechellia desatf elof Gleason, James, Wicker-Thomas, Ritchie submitted

19 Desat1, Desat2, DesatF 1 60 Desat1 MPPNAQAGAQ SISDSLIAAA SAAADAGQSP TKLQEDSTGV LFECDVETTD GGLVKDITVM Desat MAPYSRIYH QDKSSRETGV LFEDDAQTVD SDLTTDRFQL DesatF M PRNTKDTTGV LYESDVETVD GGLSTELSNR Desat1 KKAEKRLLKL VWRNIIAFGY LHLAALYGAY LMVTSAKWQT CILAYFLYVI SGLGITAGAH Desat2 KRAEKRRLPL VWRNIILFAL VHLAALYGLH SIFTRAKLAT TLFAAGLYII GMLGVTAGAH DesatF KTTDGSKLEL VWFNIVLFVI LHISSLYGVW LLFTSATWTT VVLFWPTVVV TILGVSGGAH Desat1 RLWAHRSYKA KWPLRVILVI FNTIAFQDAA YHWARDHRVH HKYSETDADP HNATRGFFFS Desat2 RLWAHRTYKA KWPLRLLLVI FNTIAFQDAV YHWARDHRVH HKYSETDADP HNATRGFFFS DesatF RLWAHRTFKA NTPLKLIFLF LNTLAFQDAV YYWARDHRVH HKYTETDADP YNSQRGWFFA Desat1 HVGWLLCKKH PEVKAKGKGV DLSDLRADPI LMFQKKYYMI LMPIACFIIP TVVPMYAWGE Desat2 HVGWLLCKKH PDIKEKGRGL DLSDLRADPI LMFQRKHYYI LMPLACFVLP TVIPMVYWNE DesatF HIGWLCCRKH PEVVEKGKQI DLSDLEADPL IMFQKKYYLL LMPIICFVLP TVLPMYLWGE Desat1 SFMNAWFVAT MFRWCFILNV TWLVNSAAHK FGGRPYDKFI NPSENISVAI LAFGEGWHNY Desat2 TLASSWFVAT MFRWCFQLNM TWLVNSAAHK FGNRPYDKTM NPTQNAFVSA FTFGEGWHNY DesatF SLNVSWHVMA LLRWCLSLNL IWTVNSSAHM HGMRPYDKNI CSVDQGFLIF FRVGEGYHNY Desat1 HHVFPWDYKT AEFGKYSLNF TTAFIDFFAK IGWAYDLKTV STDIIKKRVK RTGDGTHATW Desat2 HHAFPWDYKT AEWGCYSLNI TTAFIDLFAK IGWAYDLKTV APDVIQRRVL RTGDGSHELW DesatF HHVFPWDYKS AELGKYSQDV TTKFIEFMAY LGWAYDLKSV SLDLVKQRVQ RSGDGSHPVW 361 Desat1 GWGDVDQPKE EIEDAVITHK KSE Desat2 GWGDKDLTAE DARNVLLVDK SR DesatF GWGDKDQLKE DVGVTTITHQ RNEK

20 Genetic studies (transgenic flies) Oeno-Gal4 X UAS RNAi Tissue-specific specific Promoter gal 4 Oeno-Gal4 GAL4 UAS desatf-rnai RNAi transcription in oenocytes Degradation of desatf mrna

desatf RNAi knock-down in females OK72-C OK72-RNAi Size (kb) desat1 desatf desat1 desatf 2 Time (min) 10 * * 10 10 80 8 8 8 60 6 6 ** 6 4 4 4 40 2 2 2 P Percentages 20 0 0 0 0 * * 1 0.

21 desatf RNAi knock-down in females OK72-C OK72-RNAi Size (kb) desat1 desatf desat1 desatf 2 Time (min) 10 * * ** P Percentages * * Courtship Copulation Copulation Courtship latency latency attempts Index Wild-type male Mutant female Comportemental t test t Percentag ges *** *** *** OK72-RNAi Control OK72-RNAi *** Have less pheromones (dienes) more monoenes *** Are less courted by wild-type males 0 7-T 7-P 7-H 7,11-HD 7,11-ND saturated Chertemps,, Duportets,, Labeur, Ueyama, Wicker-Thomas Insect Mol. Biol.

22 After desatf knockdown in females (D. melanogaster) No more expression of desatf Very little production of dienes Attractivity of RNAi females decreased toward wild-type males What is the effect of desatf expression in D. simulans?

23 Evolution of pheromones D. melanogaster (Dmel) diene synthesis attractivity of females +/+ 37% dienes Df(3L)desatF 14% dienes D. simulans(dsim) desatf non transcribed no dienes F0 Df(3L)desatF x sim + sim Dmel / Dsim F1 Df(3L)desatF sim + sim desatf allele 0 1 Hydrocarbons 0% diene 14% dienes

24 100 Courtship of Dsim males toward hybrid females Dsim ** *** Percentag ges Time (min) Number 50 Dmel-/Dsim l/d Males performing courtship Males performing copulation Courtship latency Copulation latency Copulation attempts With Dmel/Dsim hybrids: little courtship, no copulation attempts, no copulation

25 100 Percentag ges 50 Courtship of Dsim males toward hybrid females Dsim Dmel Df /Dsim Dmel/Dsim Time (min) ** * Numbe er Males perf forming cou urtship Males perf forming cop pulation 0 Cou urtship lat tency Copu ulation lat tency With Dmel/Dsim hybrids: little courtship, no copulation attempts, no copulation With Dmel/Dsim hybrids without desatf: normal courtship, copulation attempts, copulation 0 Copu ulation atte empts Legendre, Miao, Da Lage, Wicker-Thomas, 2008, IBMB

26 Partial conclusion DesatF is the only desaturase involved in the synthesis of dienes (second double-bond) Dmel/Dsim females are much more attractive ti to Dsim males if they have no functional desatf allele (no dienic hydrocarbons) Dienes and desatf could play a role in reproductive isolation between Dsim and Dmel How has the gene evolved?

27 Alignment of DesatF deduced sequences 1 90 meltai MPRNTKDTTG VLYESDVETV DGGLSTELSN RKTTDGSKLE LVWFNIVLFV ILHISSLYGV WLLFTSATWT TVVLFWPTVV VTILGVSGGA melcs MPRNTKDTTG VLYESDVETV DGGLSTELSN RKTTDGSKLE LVWFNIVLFV ILHISSLYGV WLLFTSATWT TVVLFWPTVV VTILGVSGGA melweb MPRNTKDTTG VLYESDVETV DGGLSTELSN RKTTDGSKLE LVWVNIVLFV ILHISSLYGV WLLFTSATWT TVVLFWPTVV VTILGVSGGA simweb MPPNTKDKTV VLYESDAQTV DGGLSTELSN PKTTDGRKMK LVWFNIVLFL ILHISSLYGV WLFFTSATWT TFLLLWPAVV VTILGVSGGA simyb MPRNTKDTTG VLYESDAQTV DSGLSTELSN PKTTDGRKMK LVWFNIVLFL ILHISSLYGV WLFFTSATWT TFLLLWPAVV VTILGVSGGA simsey MPRNTKDTTG VLYESDAQTV DSGLSTDLSN PKTTDGRKLK LVWFNIVLFL ILHISSLYGV WLFFTSATWT TFLLLWPAVV VTILGVSGGA meltai HRLWAHRTFK VNTPLKLIFL FLNTLAFQDA VYYWARDHRV HHKYTETDAD PYNSQRGWFF AHIGWLCCRK HPEVVEKGKQ IDLSDLEADP melcs HRLWAHRTFK ANTPLKLIFL FLNTLAFQDA VYYWARDHRV HHKYTETDAD PYNSQRGWFF AHIGWLCCRK HPEVVEKGKQ IDLSDLEADP melweb HRLWAHRTFK ANTPLKLIFL FLNTLAFQDA VYYWARDHRV HHKYTETDAD PYNSQRGWFF AHIGWLCCRK HPEVVEKGKQ IDLSDLEADP simweb HRLWAHRTFK ANTPLKLIFL FLNTLAFQDA VYYWARDHRV HHKYTETDAD PYNSQRGWFF AHIGWLCCRK HPDVVEKGKQ IDLSDLEADP simyb HRLWAHRTFK ANTPLKLIFL FLNTLAFQDA VYYWARDHRV HHKYTETDAD PYNSQRGWFF AHIGWLCCRK HPDVVEKGKQ IDLSDLEADP simsey HGLWAHRTFK ANTPLKLIFL FLNTLAFQDA VYYWARDHRV HHKYTETDAD PYNSQRGWFF AHIGWLCCRK HPDVVEKGKQ IDLSDLEADP meltai LIMFQKKYYL LLMPIICFVL PTVLPMYLWG ESLNVSWHVM ALLRWCLSLN LIWTVNSSAH MHGMRPYDKN ICSVDQGFLI FFRVGEGYHN melcs LIMFQKKYYL LLMPIICFVL PTVLPMYLWG ESLNVSWHVM ALLRWCLSLN LIWTVNSSAH MHGMRPYDKN ICSVDQGFLI FFRVGEGYHN melweb LIMFQKKYYL LLMPIICFVL PTVLPMYLWG ESLNVSWHVM ALLRWCLSLN LIWTVNSSAH MHGMRPYDKN ICPVDQGFLI FFRVGEGYHN simweb LIMFQKKYYL LLMPIICFVL PTVVPMYLWG ESLNVSWHVM TLLRWCISLN LIWTVNSSAH MHGMRPYDKN ICPVDQSFLI FFHVGEGYHN simyb LIMFQKKYYL LLMPIICFVL PTVVPMYLWG ESLNVSWHVM TLLRWCISLN LIWTVNSSAH MHGMRPYDKN ICPVDQSFFI SFHVGEGYHN simsey LIMFQKKYYL LLMPIICFVL PTVVPMYLWG ESLNVSWHVM TLLRWCISLN LIWTVNSSAH MHGMRPYDKN ICPVDQSFLV FFHVGEGYHN meltai YHHVFPWDYK SAELGKYSQD VTTKFIEFMA YLGWAYDLKS VSLDLVKQRV QRSGDGSHPV WGWGDKDQLK EDVGVTTITH QRNEK melcs YHHVFPWDYK SAELGKYSQD VTTKFIEFMA YLGWAYDLKS VSLDLVKQRV QRSGDGSHPV WGWGDKDQLK EDVGVTTITH QRNEK melweb YHHVFPWDYK SAELGKYSQD VTTKFIEFMA YLGWAYDLKS VSLDLVKQRV QRSGDGSHPV WGWGDKDQLK EDVGVTTITH QRNEK simweb YHHVFPWDYK SAELGKYSQD VTTKFIEFMA YLGWAYDLKS VSLDSVKQRA QRTGDGSHPV WGWGDKDQLK EDVGVTTISH QRNGK simyb YHHVFPWDYK SAELGKYSQD VTTKFIEFMA YLGWAYDLKS VSLDSVKQRA QRTGDGSHPV WGWGDKDQLK EDVGVTTISH QRNEK simsey YHHVFPWDYK SAELGKYSQD VTTKFIEFMA YLGWAYDLKS VSLDSVKQRA QRTGDGSHPV WGWGVKDQLK EDVGVTTISH QRNEK

28 Alignment of promoter sequences meltai -634 CATCTACTGTCTGCACGCAGAAAAAAAATCAAACAGCCACTCAGTTTTCGATAAATTAATAAACCAAAAATAGCACACAGCTATAATTTGT-----AAAAAATAAGACTAAAATTATTCA melcs -636 CATCTACTGTCTGCACGCAGAAAAAAAATCAAACAGCCACTCAGTTTTCGATAAATTAATAAACCAAAAATAGCACACAGCTATAATTTGT-----AAAAAATAAGACTAAAATTATTCA melweb -638 CATCTACTGTCTGCACGCAGAAAAAAAATCAAACAGCCACTCAGTTTTCGATAAATTAAT AAACCAAAA ATAATTTGT-----AAAAAATAAGACTACAATTATTCA simweb -717 CATCTACTGTGTGCACGC--AGAAAAAATCAAATAGCCACTCAGTTTTCGATAAATTAATAAACCAAAAGTATAATAGAGCCATAATTTGA----AAAAAAATGAGACTACAGTTACTAA simyb -715 CATCTACTGTGTGCACGC--AGAAAAAATCAAGCAGCCACTCAGTTTTCGATAAATTAATAAACCAAAAATAGCACACAGCTATAATTTGT-----AAAAAATAAGACTAAAATTATTCA simsey -713 CATCTACTGTCTGCACGC--AGAAAAAATCAAACAGCCACTCAGTTTTCGATAAATTAATAAACCAAAAGTATAATAGAGCCATAATTTGTaaaaAAAAAAATAAGACTACAGTTACTAAC C C GC CGC G C C GCC C C G CG CC G G GCC G G C C G C meltai -519 ATTCAACTTATTA--TTC--AAAA--ATTAATAATTATATAATA AAACATTTAAAGCATTGCAGTTCTTGCTAGATACTTTTGATATTTTTGAATTGAAT----ATTGATT melcs -521 ATTCAACTTATTA--TTC--AAAA--ATTAATAATTATATAATA AAACATTTAAAGCATTGCAGTTCTTGCTAGATACTTTTGATATTTTTGAATTGAAT----ATTGATT melweb -536 ATTTAACTTATTA-TTTA---AAAC--TAAATAATTATATAATA AAACATTTAAAGCATTACAGTTCTTGCTAGATACTTTTGATATTTTTGAATTGAAT----ATTGATT simweb -603 ATGCAAATTATTAtTTTAAAAAAATTATTATAATATGTAAAAAA------AACAAACATTGGAAGTATTGCAGTTCTTGCTAGATACTTCTAATATTTTTGAACTGAATTAAATATTGAA simyb -601 ATTTAAGTTATTA-TTTAAAAAAAT-ATTAAAATATGTAAAAAAgcaaacATAAAACATTTGAAGTATTGCAGTTCTTACTAGATACTTTTAATATTTTTGAACTGAATTGAATATTGAA simsey -595 ATGTAAATTATTA-TTTAAAAAAACAATTATAATATGTAAAAAA ACAAACA--TAAAACATT----TTCTTTCTTGATACTTTTAATATTTTTGAACTGAATTGAATGTTGAA meltai -418 ATTTTAGGTAGACTGGAGGTAATGCAAAATGTTGGAAGTTATCTGATGCAAATAAAAATAGCCCCGTAAATACCAGCAAAATTTATAAAATTGTACGATTCAGAATTTGAATTGGTAAAA melcs -420 ATTTTAGGTAGACTGGAGGTAATGCAAAATGTTGGAAGTTATCTGATGCAAAT-AAAATAGCCCCGTAAATAGCAGCAAAATTTATAAAATTGTACGATTCAGAATTTGAATTGGTAAAA melweb -435 ATTTTAGGTAGACTGGAGGTAATGCAAAATGTTGGAAGTTATCTGATGCAAATAAAAATAGCCCCGTAAATACCAGCAAAATTTATAAAATTGTACGATTCAGAATTTGAATTGGTAAAA simweb -509 GATTTAAGGTGACTGGAGGTAATGGTCAAAGTAGGAAGTAATCTGATGCAAATAAAAATAACCCCGTAAATACCAG-GAAATTTATAAAATTGTACGATTCAGAAACTCATTCGATCGCC simyb -483 GATTTTAGGTGACTGGAGGTAATGGTCAAAGTATGAAGTAATCTGATGCAAATAAAAATAACCCCGTAAATACCAG-GAAATTT ATTTAGAAACTCAATCGATCGCA simsey -489 GATTTAAGGTGACTGGAGGTAATGGACAAAGTATGAAGTAATCTGATGCAAATAAAAATAACCCCGTAAATACCAG-GAAATTTATAAAATTGTGCGACTTAGAAACTCAATCGATCACA meltai CCGAAAATGCATAGCTTTCCAGGAGTGCAACAATGTATGTATATGTC--AATGCAAATTTGAATTGCATTCGTAG CA--CTGCGTAAGCAGGAACCG melcs CCGAAAATGCATAGCTTTCCAGGAGTGCAACAATGTATGTATATGTCATAATGCAAATTTGAATTGC-TTCGTAG CA--CTGCGTAAGCAGGAACCG melweb CCGAAAATGCATAGCTTTCCCTGAGTGCAACAATGTATGTATATGTCATAATGCAAATTTGAATTGCATTCGTAG CA--CTGCGTAAGCAGGAACCG simweb -370 ATTCGAATTGTTAAAACCGAAAATGCATAGCTTTCCAAGAGTGCAACAATGTATATGTCAGAATTCATGTCATTCATGAATGCCATTATTT CATAATGCGTAA-CAGGAACCG simyb -377 ATTCGAATTGGTAAAACCGAAAATGCATAGCTTTCCAAGAGTGCAAGAATCTATGTGTCAGAATTCATGTCATTCATGAATGCCATTATTTcatcctgCATAATGCGTAA-CAGGAACCG simsey -370 ATTCGAATTGTTAAAACCGAAAATGCATAGCTTTCCAAGAGTGCAACAATGTATATGTCAGAATTCATGTCATTCATGAATGCCATTATTT CATAATGCGTAA-CAGGAACCG meltai -203 AAGAAATTCAA-TTTGTTTAAGCGAATTTTTGCACTTTAATTTGTTGCATTCAAA TTTGTAATTT GTAATTmelCS -207 AAGAAATTCAAtTTTTTTTAAGCAAATTTTTGCACTTTAATTTGTTGCATTCAAA TTTGTAATTT GTAATTmelweb -220 AAGAAATTCAA-TTTGTTTAAGCGAATTTTTGCACTTTAATTTGTTGCATTCAAA TTTGTAATTT--GTTAATTTGTA------AT--TTGTAATTsimweb -258 AAGAATTTAAA-TTTTTTAAAACGAATTTTTGCATTTTATTTTGTTGCATTCAAAGCGATTCCATGCTTATGCTTCGATTGCTAATTAATGCGAACTTCTATCATTCATGGGCGTAATTA simyb -258 AAAAATTTTAA-TTTTTGAAAGCGAATTTTCGCATTTTATTTTGTTGCATTCAAAGCGATTCCATGCTTATGCTTCGATTGCTAATTAATGCGAACTTCTATCATTCATGGGCGTAATTA simsey -258 AAGAATTTAAA-TTTTTTAATGCGAATTTTCGCATTTTATTTTGTTGCATTCAAAGCGATTCCATGCTTATGCTTCCATTGCTAATTAATGCGAACTTCTATCATTCATGGGCGTAATTA meltai TGCAAATAGCGAGCGCTTGAAGGTCATCGATTTGAATTGCACTCCGACGTTATGGGTTCAACGTATCATGTTTATATGTATCTGATAAATAGAAGCCCAGTCGAAAAGCAGAAACA melcs TTGCAAATAGCGAGCGCTTGAAGCGCATAGATTTGAATTGCACTCCGACGTTATGGGTTCAACGTATCATGTTTATATGTATCTGATAAATAGAAGCCCAGTCGAAAAGCAGAAACT melweb TGCAAATAGCGAGCGCTTGAAGCGCATAGATTTGAATTGCACTCCGACGTTATGGGTTCAACGTATCATGTTTATATGTATCTGATAAATAGAAGCCCAGTCGAAAAGCAGAAACT simweb -139 GCGATGCAGACAGCGAGCGCTTGTAGGTCATGAATTTGAATTCCACTCCGACATTATGGGTTCTACGAATTATTTTTATACGTATCTGATAAATAGAAGCCCAGTCCAAAAGCAGAAACA simyb -139 GCGATGCAGACAGCGAGCGCTTGTAGGTCATGAATTTGAATTCCACTCCGACATTATGGGTTCTACGAATTATTTTTATACGTATCTGATAAATAGAAGCCCAGTCCAAAAGCAGAAGCA simsey -139 GCGATGCAGACAGCGAGCGCTTGTAGGTCATGAATTTGAATTCCACTCCGACATTATGGGTTCTACGAATTATTTTTATACGTATCTGATAAATAGAAGCCCAGTCCAAAAGCAGAAGCA meltai -19 GTATCAAACGTAAAGCACC ATGCCACGCAATACCAAAGACACCACCGGAGTGCTTTACGA melcs -19 GTATCAAACGTAAAGCACC ATGCCACGCAATACCAAAGACACCACCGGAGTGCTTTACGA melweb -19 GTATCAAACGTAAAGCACC ATGCCACGCAATACCAAAGACACCACCGGAGTGCTTTACGA simweb -19 GTATCAAACATAGAGCAAC ATGCCACCCAATACCAAAGACAAAACCGTAGTGCTTTACGA

29 Percentages of identities (gaps) between sequences mel CS mel Tai mel mel mel sim sim sim CS Ti Tai web Sey YB web 97.6 (2.2) 97.6 (2.2) 93.8 (5.9) 94.5 (4.9) 74.8 (24.7) 76.4 (23.1) 75.8 (23.3) 77.2 (21.9) 76.8 (22.3) 78.4 (20.8) mel web (5.9) (4.9) (23.2) 2) (21.1) 1) (20.9) sim Sey sim YB 74.8 (24.7) 75.8 (23.3) 76.4 (23.1) 77.2 (21.9) 76.3 (23.2) 77.8 (21.1) 90.1 (8.6) 90.1 (8.6) sim web (22.3) (20.8) (20.9) (6.6) (8.6) 92.7 (6.6) 90.7 (8.6) Without any selection pressure Divergence mel/sim : 4-8% desatf promoter mel/sim:22-25%

30 Conclusions desatf has evolved very rapidly between D. simulans and D. melanogaster It has a large effect on D. simulans courtship It may have played a role in sexual isolation leading to speciation

31 Evolution of desatf desatf has evolved very rapidly between Drosophila species. Its evolution in Drosophila is under study (Maria Keays, Mike Ritchie Lab)

32 ACKNOWLEDGMENTS NAMC (Orsay) Thomas Chertemps Carole Labeur Renaud Dallerac Arièle Legendre Line Duportets Xuexia Miao (China) Jean-Marc Jallon Morio Ueyama (Japan) LEGS (Gif) Ilhem Guenachi Mike Ritchie s lab

33 Thank you for your attention

Genetic Contributions of desatf and elof to Courtship Mating Behavior and Cuticular Hydrocarbon Production in Drosophila simulans and D.

Genetic Contributions of desatf and elof to Courtship Mating Behavior and Cuticular Hydrocarbon Production in Drosophila simulans and D. sechellia BY Jennifer L. Hackett Submitted to the graduate degree