SUPPLEMENTARY MATERIAL

Transcription

1 /ZO13105_AC CSIRO 2014 Australian Journal of Zoology 2014, 62(3), SUPPLEMENTARY MATERIAL The koala immunological toolkit: sequence identification and comparison of key markers of the koala (Phascolarctos cinereus) immune response Identification of koala immunological markers Katrina Morris A, Peter J. Prentis B, Denis O Meally A, Ana Pavasovic C, Alyce Taylor Brown D, Peter Timms D,E, Katherine Belov A and Adam Polkinghorne D,E,F A Faculty of Veterinary Science, University of Sydney, RMC Gunn, B19, Sydney, NSW 2006, Australia. B School of Earth, Environmental and Biological Sciences, Queensland University of Technology, 2 George Street, Brisbane, Qld 4001, Australia. C School of Biomedical Sciences, Queensland University of Technology, 2 George Street, Brisbane, Qld 4001, Australia. D Institute of Health and Biomedical Innovation, Queensland University of Technology, 60 Musk Avenue, Kelvin Grove, Qld 4059, Australia. E Faculty of Science, Health, Education and Engineering, University of the Sunshine Coast, 90 Sippy Downs Drive, Sippy Downs, Qld 4556, Australia. F Corresponding author. a.polkinghorne@qut.edu.au

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3 Supplementary Figure 1: CD4 amino acid alignment in koala (Phascolarctos cinereus), wallaby (Macropus eugenii; EU152103), devil (Sarcophilus harrisii; KF640217), opossum (Monodelphis domestica; NM_ ) and human (Homo sapiens; M12807). Dots indicate identity to koala sequence. The four extracellular domains, transmembrane (TM) region and cytoplasmic tail (CYT) were predicted through alignment to marsupial sequences. Conserved extracellular cysteine residues, a cysteine residue unique to wallaby and devil and a cysteine residue replaced by a tryptophan in marsupials are indicated by stars, a square and a diamond below the residues, respectively.

4 Supplementary Figure 2: CD8α amino acid alignment in koala (Phascolarctos cinereus), wallaby (Macropus eugenii; EF490599), devil (Sarcophilus harrisii; KF640218), opossum (Monodelphis domestica; NM_ ) and human (Homo sapiens; M12828). Dots indicate identity to koala sequence. The IgSfV domain, hinge segment, transmembrane (TM) region and cytoplasmic tail (CYT) were predicted by alignment to marsupial sequences. Conserved extracellular cysteine residues are indicated by stars below the residues.

5 Supplementary Figure 3: CD8β amino acid alignment in koala (Phascolarctos cinereus), wallaby (Macropus eugenii; EU152105), devil (Sarcophilus harrisii; ENSSHAT ), opossum (Monodelphis domestica; NM_ ) and human (Homo sapiens; BC100911). Dots indicate identity to koala sequence. The IgSfV domain, hinge segment, transmembrane (TM) region and cytoplasmic tail (CYT) were predicted by alignment to marsupial sequences. Conserved extracellular cysteine residues are indicated by stars below the residues.

6 Supplementary Figure 4: IL-4 alignment between alignment in koala (Phascolarctos cinereus), wallaby (Macropus eugenii; ADG01643), devil (Sarcophilus harrisii; XP_ ), opossum (Monodelphis domestica; XP_ ) and human (Homo sapiens; NP_000580). Dots indicate identity to koala sequence. The IL4 family signature motif is boxed.

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8 Supplementary Figure 5: IgA heavy chain constant region alignment in koala (Phascolarctos cinereus), possum (Trichosurus vulpecula; AAD41690), opossum (Monodelphis domestica; AAC48835) and human (Homo sapiens; AAC82528, AAB59396). Dots indicate identity to koala sequence. The location of the three heavy chain constant (CH1-CH3) and hinge (H) regions are indicated above the sequence. Location of 8 cysteine residues involved in formation of intra-chain disulphide bridges in human IgA are indicated by stars below the residue.

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10 Supplementary Figure 6: IgG heavy chain constant region alignment in koala (Phascolarctos cinereus), possum (Trichosurus vulpecula; AAD55590), opossum (Monodelphis domestica; AAC78139) and human (Homo sapiens; AAC82527, AAN76044, AA67886, CAC20457). The location of the three heavy chain constant (CH1-CH3) and hinge (H) regions are indicated above the sequence. Location of 6 cysteine residues involved in formation of intra-chain disulphide bridges in human IgG are indicated by stars below the residue.

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12 Supplementary Figure 7: IgM heavy chain constant region alignment in koala (Phascolarctos cinereus), possum (Trichosurus vulpecula; AAF80358), opossum (Monodelphis domestica; AAC48834) and human (Homo sapiens; CAC20458). Dots indicate identity to koala sequence. The location of the four heavy chain constant (CH1-CH4) regions are indicated above the sequence. Location of 8 cysteine residues involved in formation of intra-chain disulphide bridges in human IgM are indicated by stars below the residue

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14 Supplementary Figure 8: IgE heavy chain constant region alignment in koala (Phascolarctos cinereus), possum (Trichosurus vulpecula; AAF80357), opossum (Monodelphis domestica; AAC78140) and human (Homo sapiens; AAB59395). Dots indicate identity to koala sequence. The location of the four heavy chain constant (CH1-CH4) regions are indicated above the sequence. Location of 8 cysteine residues involved in formation of intra-chain disulphide bridges in human IgE are indicated by stars below the residue.

15 Supplementary Figure 9: IL-6 alignment in koala (Phascolarctos cinereus), devil (Sarcophilus harrisii; ENSSHAT ), opossum (Monodelphis domestica; ENSMODP ) and human (Homo sapiens; CAG29292). Dots indicate identity to koala sequence. Four conserved cysteine residues are indicated by a star below the residue. The IL-6 family signature motif is boxed.

16 Supplementary Figure 10: IL-12A alignment in koala (Phascolarctos cinereus), opossum (Wong et al., 2006) and human (Homo sapiens; NP_000873). Dots indicate identity to koala sequence.

17 Supplementary Figure 11: IL-12B alignment in koala (Phascolarctos cinereus), devil (Sarcophilus harrisii; XP_ ) and human (Homo sapiens; NP_002178). Dots indicate identity to koala sequence.

18 Supplementary Figure 12: IL-1β alignment in koala (Phascolarctos cinereus), wallaby (Macropus eugenii; ACU65957), possum (Trichosurus vulpecula; AAD21871) and human (Homo sapiens; AAA59134). Dots indicate identity to koala sequence. The propeptide sequence is underlined. A conserved histidine is indicated by a star below the residue. The IL-1 family signature motif is boxed.