<p> 1</p><p>Supplementary Material</p><p>IpFPPS MF-SMMACRNRPCREWLKEFRRNISKTSTDKNSDAIIRSQDKMQHASKTHEVAFEKNLSY 59 DjFPPS MF-SMKLCRNRSCREFLREARRTISKTSTDKNSGAISRAPDHKLN------VESDSTGSY 53 TfFPPS MFPSVKICVNRTSRDILRKFQRHISKTSSVPNSDAISRQQDNTLKVN-HDLEAYNRNKSS 59 CfFPPS1 MF-STTRSLEKIVQSYKNELRRQISKTTSVTNSDAMAPRLDQPANKT----PTNEETGLP 55 MpFPPS1 MY------KTLTTFTRALSRRTAFSLCPAVAAG------27 *: . * :*: :: . *: </p><p>IpFPPS TRWMKQMQHNNIRALSTIQQTLVRPQQSSSLASKEQSRDFMAQFPDIIRELTEVGRNQEL 119 DjFPPS SRWKKQMHHNNIRALSTIQQSMIRPVQSSALVTKEQSRDFMALFPDLVRELTEVGKSQEL 113 TfFPPS IKWAKLSRHNNIRALSTIQTKVKPPVTSTPLVSKEESREFMAIFPDIVRDLTDAGRHTDI 119 CfFPPS1 KKLLKLQKYH--RFLSTLTPQQMPLAARGLAVSKDQSREFMACFPDIVRDLTETSKHVDV 113 MpFPPS1 ------RENHFRSMSVILPPPVTSVITGALVSKDELGNFMAVFPDIVKDLTDTSLQLNK 80 : : * :*.: .:*:: :*** ***::::**:.. : </p><p>IpFPPS PEVMKRYARVLQYNTPNGKKTRGLITISTYKMLEDPAKITTENLKKAGILGWCIEMLHTY 179 DjFPPS HDVMRRFARVLQYNTPTGKKNRGLIILSTYRMLEDPEKLTPENIRLASILGWCAEMVHAY 173 TfFPPS PEVTKRFAKVLQYNVPTGKKTRGLSCVTAYKILEKPENLTPENVKLANILGWCIELLRGF 179 CfFPPS1 PEASRWLAKLLQYNVPNGKKNRGLATVLAYKMLEKPENLTPENVHLANVMGWCTEMFHTH 173 MpFPPS1 PDVTKWLENLLQYNVPGGKNNRGLILVSSFKMLSSPSYLTDENLRLSYILGWCVEILQAY 140 :. : .:****.* **:.*** : ::::*..* :* **:: : ::*** *:.: .</p><p>IpFPPS FLIIDDIIDHSDTRRGAICWYRQPGIGLTAVYDAVMMENGVYLLLKRHFKDHPMYTNIIE 239 DjFPPS VLILDDIMDGSETRRGALCWFRQSGIGLTAVNDAVMIENAVYLLIKRHLKDHPMYVPLME 233 TfFPPS ELIVDDIVDNAETRRNAPCWFRKDNVGYFAIGDSILLESSVYSILRKYFSTLPCYVPMME 239 CfFPPS1 QLLLNDIMEGTEMRRGAPAWHRRPDVGLSSINDAILVQSAMYSTLKKHFHAKPYYKNVIE 233 MpFPPS1 QLVMDDLMDNAITRRGRPCWYRHNDVGLMAVNDGILLEQAIYQLLKKYFKDKPYYTHILE 200 *:::*::: : **. .*.*: .:* :: *.::::..:* ::::: * * ::*</p><p>IpFPPS LFHDMALKTSLGQSLDTMCLNDDGKPKLDIHNE-QDTSIVNYKTSYYSFYLPVAAAMYLL 298 DjFPPS LFHEGNLKTTLGQSLDAMCLDTNGKPKLDMFTMSRYTSIVKYKTAFYSFQMPVAIAMYLA 293 TfFPPS LVHDVTLKTAMGKSLEGLIMQ-GGKPNLDKFTMKNYNLMTKYKVGYYTFQLPVALAMYFA 298 CfFPPS1 MFNEMLLKCSTGQFLEKAMAK-TDKPDLSQFTMEKYLTIAKYKTAYHTFQMPVGLALLMS 292 MpFPPS1 LFYDVTMKSAMGQCLDMLTAKSFKSKKLEKYTMENYKAIVKYKTAYYSFVLPVCLAMRMT 260 :. : :* : *: *: . . .*. .. . :.:**..:::* :** *: : </p><p>IpFPPS GMDDPEQHRQARTILLDMGQFFQIQDDFLDCFGDPNVTGKLGTDIQDGKCSWLAVMALQR 358 DjFPPS GMSDEEQHRQAKTILMEMGQFFQIQDDFLDCFGDPTVTGKVGTDIQDGKCSWLAVVALQR 353 TfFPPS NMFDAEQHRQAKTILLEMGQFLQIQTDFLDCFGDP--AGKIGNDIQAGRCSWLAIVALQR 356 CfFPPS1 GVDDLETHRQAKTILLEMGQFFQIQDDFLDCFGDPAVTGKNGSDIQDGKCTWLAVVALQR 352 MpFPPS1 NVNDQEIFRQAKVILLDMGQFFQIQDDYLDCYGNPEITGKIGTDIEDGKCSWLAVKALQK 320 .: * * .***:.**::****:*** *:***:*:* :** *.**: *:*:***: ***:</p><p>IpFPPS STPAQRKIMEEHYGRSEPESVAIIKQLYEDLTLPNTYAIYEEESFNIIKTHIQQISKDCV 418 DjFPPS ASPAQRKIMEEHYGRPEPESIARIKNLYVDLCLPNTYAIYEEESFNIIKTHIQQISKGLR 413 TfFPPS ANPAQRKLMEDYYGMPGAQAGQIIKMLYEELSLPATYAAYEEESYNIINTHIQQISKGLP 416 CfFPPS1 ASPAQRHVLEEHYGSSKPEDVEKIKELYETLQLPHTYSVYEDATYDLLRTQIQQVTRGLP 412 MpFPPS1 VTTEQKKIMEDNYGIDDQFNVAVIKDLYQQLKLQNTFLLYEEESYNLIRTCIQNFSPGLS 380 .. *::::*: ** ** ** * * *: **: :::::.* **:.: . </p><p>IpFPPS T-IFSKHYGEAFKRES--- 433 DjFPPS HDLFLKIMENIYKREC--- 429 TfFPPS HELFFTLMEKLYRINEYDC 435 CfFPPS1 HDLFFKILDNIFRRNI--- 428 MpFPPS1 QDMFFKLLEKIYKRTL--- 396 :* . : :: </p><p>Supplementary Figure 1. ClustalW alignment of TfFPPS with two other coleopteran FPPSs [Ips pini FPPS (IpFPPS); Dendroctonus jeffreyi FPPS (DjFPPS), as well as with a lepidopteran FPPS1 [Choristoneura fumiferana FPPS1 (CfFPPS1)] and an aphid FPPS1 [Myzus persicae FPPS1 (MpFPPS1)]. The sequence shaded in gray is the first aspartate-rich motif (FARM). 2</p><p>↓ FARM DmFPPS FFIISDDVMD ApFPPS YQLVLDDIMD MpFPPS YQLVLDDIMD TfFPPS FELIVDDIVD</p><p>Supplementary Figure 2. Amino acid alignment of the FARM region (DDxxD) of TfFPPS with the corresponding residues in Drosophila melanogaster FPPS (DmFPPS) and two aphid FPPSs [Acyrthosiphon pisum FPPS (ApFPPS) and Myzus persicae FPPS (MpFPPS)]. The arrow points to the residue at position -4 relative to the FARM, which is known to play an important role on controlling product chain length in short-chain prenyltransferases such as FPPS. In most eukaryotes, this residue is aromatic (F/Y) but a glutamine (Q) substitutes for F/Y in aphid FPPSs, which can produce both C10 (geranyl diphosphate) and C15 (FPP) products (Vandermoten et al. 2008). Site-directed mutagenesis experiments have shown that the replacement of F/Y by Q was responsible for the dual GPP/FPP synthase activity of aphid FPPSs (Vandermoten et al. 2009). The Glu residue at the same position in TfFPPS is expected to have a similar impact on product chain length; it is therefore possible that this enzyme displays the same dual activity observed for aphid FPPS. However, the recombinant TfFPPS produced using a procedure used for other FPPSs (Cusson et al., 2006; Sen et al., 2007; Vandermoten et al., 2008) did not display prenyltransferase activity under in vitro conditions.</p><p>CUSSON, M., BÉLIVEAU, C., SEN, S.E., VANDERMOTEN, S., RUTLEDGE, R.J., STEWART, D., FRANCIS, F., HAUBRUGE, É., REHSE, P., HUGGINS, D.J., DOWLING, A.P.G., and GRANT, G.H. 2006. Characterization and tissue-specific expression of two lepidopteran farnesyl diphosphate synthase homologs: implications for the biosynthesis of ethyl-substituted juvenile hormones. Proteins 65:742-758.</p><p>SEN, S.E., TROBAUGH, C., BÉLIVEAU, C., RICHARD, T. and CUSSON, M. 2007. Cloning, expression and characterization of an insect farnesyl diphosphate synthase. Insect Biochem. Mol. Biol. 37:1198-1206.</p><p>VANDERMOTEN, S., CHARLOTEAUX, B., SANTINI,, S., SEN, S.E., BÉLIVEAU, C., VANDENBOL, M., FRANCIS, F., BRASSEUR, R., CUSSON, M., and HAUBRUGE, É. 2008. Characterization of a novel aphid prenyltransferase displaying dual geranyl/farnesyl diphosphate synthase activity. FEBS Letters 582:1928-1934. 3</p><p>VANDERMOTEN, S., SANTINI, S., HAYBRUGE, É., FRANCIS, F., BRASSEUR, R., CUSSON, M., and CHARLOTEAUX, B. 2009. Structural features conferring dual GPP/FPP synthase activity to an aphid prenyltransferase. Insect Biochem. Mol. Biol. 39:707-716.</p>