Figure S6.
Actin cytoskeletal drugs bind sequences that are conserved in Naegleria proteins.(A) The alignment shown in Fig. S5 was used to calculate the percent identity of actin protein sequences from representative eukaryotes. Values indicating higher conservation are highlighted in green, and values indicating less conservation are highlighted in lilac. Arp1 from rabbit was included for comparison to an actin-related protein (shown in gray). (B) Drug binding sites on mammalian actin (Faulstich et al., 1993; Morton et al., 2000; Nair et al., 2008; Pospich et al., 2017) are shown (gray) in comparison to Giardia actin and Naegleria actin. Identical residues are shown in green, non-identical residues are in lilac, and the numbers above each column indicate the location of the residue in the rabbit actin sequence. Giardia actin is shown as an example due to its low percent identity to mammalian actin, as well as the documented ineffectiveness of actin inhibitors (Paredez et al., 2011). A question mark indicates unknown information. (C) Residues of Arp2 and Arp3 that exist within 5 Å of the CK-666 binding site on Bos taurus (gray) Arp2/3 complex (PDB 3UKR; Baggett et al., 2012) were compared between model organisms and Naegleria. Identical residues are shown in green, non-identical redidues are highlighted in lilac. (D) FH2 domains from six SMIFH2-sensitive formins in diverse model organisms (including Mus musculus and Arabidopsis thaliana) were used to generate an MSA, and the percent identity was calculated in comparison to each other and compared with the FH2 domains of Naegleria’s 14 formins. Higher percent identity values are shown in green, lower values are shown in lilac. (E) SMIFH2-sensitive formins shown in D were used to calculate an FH2 domain consensus sequence (gray). Positions with at least 75% identity among SMIFH2-sensitive FH2 domains are shown, with positions of Naegleria FH2 domains that are identical to the consensus shown in green. The total percent identity to the consensus sequence is shown to the right, with higher percent identities in green, and lower percent identities in lilac. All accession numbers for the sequences used to generate this figure are listed in Table S3.

Actin cytoskeletal drugs bind sequences that are conserved in Naegleria proteins. (A) The alignment shown in Fig. S5 was used to calculate the percent identity of actin protein sequences from representative eukaryotes. Values indicating higher conservation are highlighted in green, and values indicating less conservation are highlighted in lilac. Arp1 from rabbit was included for comparison to an actin-related protein (shown in gray). (B) Drug binding sites on mammalian actin (Faulstich et al., 1993; Morton et al., 2000; Nair et al., 2008; Pospich et al., 2017) are shown (gray) in comparison to Giardia actin and Naegleria actin. Identical residues are shown in green, non-identical residues are in lilac, and the numbers above each column indicate the location of the residue in the rabbit actin sequence. Giardia actin is shown as an example due to its low percent identity to mammalian actin, as well as the documented ineffectiveness of actin inhibitors (Paredez et al., 2011). A question mark indicates unknown information. (C) Residues of Arp2 and Arp3 that exist within 5 Å of the CK-666 binding site on Bos taurus (gray) Arp2/3 complex (PDB 3UKR; Baggett et al., 2012) were compared between model organisms and Naegleria. Identical residues are shown in green, non-identical redidues are highlighted in lilac. (D) FH2 domains from six SMIFH2-sensitive formins in diverse model organisms (including Mus musculus and Arabidopsis thaliana) were used to generate an MSA, and the percent identity was calculated in comparison to each other and compared with the FH2 domains of Naegleria’s 14 formins. Higher percent identity values are shown in green, lower values are shown in lilac. (E) SMIFH2-sensitive formins shown in D were used to calculate an FH2 domain consensus sequence (gray). Positions with at least 75% identity among SMIFH2-sensitive FH2 domains are shown, with positions of Naegleria FH2 domains that are identical to the consensus shown in green. The total percent identity to the consensus sequence is shown to the right, with higher percent identities in green, and lower percent identities in lilac. All accession numbers for the sequences used to generate this figure are listed in Table S3.

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