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Supporting Information Supporting Information Kamikawa et al. 10.1073/pnas.0711084105 Mastigamoeba balamuthi AmoebozoansAmoebozoans Acanthamoeba castellanii Porphyra yezoensis Red algae 98 Cyanidioschyzon merolae Dictyostelium discoideum AAmoebozoansmoebozoans 83 Echinostelium arboreum Corallochytrium limacisporum Corallochytrium Capsaspora owczarzaki 87 IchthyophonidsIcchthyophonids Ichthyophonus irregularis Monosiga brevicolis Choanoflagellate Stylonychia lemnae CiliateCiliate Euglena gracilis Euglenozoans Trypanosoma brucei Arabidops thaliana SStreptophytestreptophytes 100 Nicotiana tabacum Acrasis rosea Heteroloboseans 0.95 Naegleria gruberi 35 Mallomonas rasilis 0.99 Heterosigma akashiwo 32 Phytophtora capsici StramenopilesStramenopiles 1.00 Phaeodactylum tricornutum 0.92 44 Thalassiosiraosira pseudonanapsseudonana 19 Plasmodium knowlesi Apicomplexa 99 Cryptosporidium parvum Reclinomonas americana JakobidJakobid Cyanophora paradoxa Glaucophyte Malawimonas jakobiformis MalawimonasMalawwimonas Trimastix pyriformis Trimastix 79 Streblomastix strix OxymonadOxymonad Blastocystis hominis Stramenopile Entamoeba histolytica EntamoebaEntamoebaa Teterahymena pyriformis Ciliates Euplotes crassus Giardia lamblia DiplomonadsDiplomonads 100 Spironucleusironucleus vvortensorrtens Tritrichomonas foetus Prabasalids 100 Trichomonas vaginalis 0.1 substitutions/site Fig. S1. Maximum-likelihood EF-1␣ phylogeny estimated by IQPNNI v.3.1. The diatom sequences are highlighted in red. The ML bootstrap values and Bayesian posterior probabilities for the nodes that are significant for the origin of the diatom homologues are shown. For other nodes, only ML bootstrap values Ն70% are indicated. Bayesian posterior probabilities Ն0.95 calculated by MrBayes v. 3.1.1 are also highlighted by dots. Kamikawa et al. www.pnas.org/cgi/content/short/0711084105 1of4 C P A /TKK motif B K indel P K indel /T KK P Fig. S2. P/TKK modtif and K indel. (A and B) /TKK modtif (A) and K indel (B) in the primary structure. Yeast EF-1␣, Thalassiosira (Thapse) EFL, and Ligulodinium (Linpol) EFL sequences are aligned. The motifs are highlighted in yellow. (C) The two motifs in the putative EF-1␣/EFL tertiary structure. The EFL motifs are mapped on yeast EF-1␣ tertiary sturcture (PDB ID code 1IJF). The putative residues that are responsible for elongation factor functions in EF-1␣ (9) are highlighted in pink. The residues that correspond to the EFL motifs are highlighted in yellow. Kamikawa et al. www.pnas.org/cgi/content/short/0711084105 2of4 Table S1. Clones obtained in the initial RT-PCR survey Species EFL EF-1␣ eRF3 HBS1 Others* Total Centrales Chaetoceras dydimus 50 4 0 4 13 Detonula confervacea 10 1 0 0 0 11 Ditylum brightwellii 11 0 0 0 0 11 Skeletonema costatum 12 0 0 2 0 14 Pennales Asterionella glacialis 41 8 0 2 15 Achnanthes kawaitensis 12 1 0 0 0 13 Cylindorotheca closterium 70 3 0 2 12 Thalassionema nitzschioides 81 0 0 0 9 eRF3, eukaryotic release factor 3; HBS1, hsp70 subfamily B suppressor 1. *Sequences that do not belong to eukaryotic EF-1␣ superfamily. Kamikawa et al. www.pnas.org/cgi/content/short/0711084105 3of4 Table S2. Primers used for the RT real-time PCR assay Primers Target Annealing temp., Gene species Direction Sequence deg. C EF-1␣ Tp Forward 5’-CCACCACAACTGGGCATCTA-3’ 55 Reverse 5’-ACTACTGTGAAATCATACTTTGG-3’ Pt Forward 5’-CACTACTGGTCATTTGATC-3’ 52 Reverse 5’-ATGACGGTAAAGGAGTATTTGG-3’ EFL Tp Forward 5’-GGACACTTGATCTTCAAGC-3’ 55 Reverse 5’-GACAATGGTGTAGTGGTACTTTTC-3’ ␣-tubulin Tp Forward 5’-CAAGTGTGATCCTCGTCACGG-3’ 60 Reverse 5’-GCAACCATGCACACGGCACG-3’ Pt Forward 5’-CAAGTGTGATCCTCGTCACGG-3’ 60 Reverse 5’-GCAACCATGCACACGGCACG-3’ Tp, Thalassiosila psuedonana; Pt, Phaeodactylum tricornutum. Kamikawa et al. www.pnas.org/cgi/content/short/0711084105 4of4.
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