657942.fig.001a
(a)
657942.fig.001b
(b)
657942.fig.001c
(c)
657942.fig.001d
(d)
657942.fig.001e
(e)
Figure 1: Structural features of EhADH112 and generation of ANeoBro1 trophozoites. (a) Structural features of representative Bro1 domain-containing proteins and EhADH112- and EhADH112-like proteins: Bro1 domain (squares containing diagonal lines), coiled coil regions (ellipses), proline-rich tracts (white squares), and adherence region (gray square). Hs: Homo sapiens; Sc: Saccharomyces cerevisiae; Eh: Entamoeba histolytica. Numbers: amino acid number of each protein. (b-c) Spatial conformation of EhADH112 and its Bro1 domain. (b) Ribbon representation for predicted EhADH112 tertiary and human ALIX crystallized structures and (c) Bro1 domains from EhADH112, ALIX and yeast BRO1 proteins. N: amino terminus. C: carboxy terminus. TPR: tetratricopeptide repeat. (d) Schematic depiction of the pNeoBro1FLAG plasmid. K: KpnI restriction site. B: BamH1 restriction site. (e) Transcripts obtained by RT-PCR assays using oligonucleotides for the NeoR gene or Bro1FLAG sequence and cDNAs synthesized from nontransfected clones A (lane 2), ANeo (lane 3), and ANeoBro1 (lane 4). Lane 1 corresponds to the reaction mixture without reverse transcriptase, using total RNA from ANeoBro1.