Figure 6

New gene model codes for a canonical GCS1-HAP2 protein, EtHAP2. (a) (i) Upregulated gametocyte transcripts, ETH_00017050 and ETH_00017055, were originally predicted as separate, tandemly positioned genes. Exons (yellow) are separated by introns (solid black line), while the intergenic regions are indicated by a broken black line. The position of the genomic sequence coding for a putative GCS1-HAP2 domain is indicated by a green box. (ii) Gametocyte RNA Seq reads were mapped to the E. tenella genome in either a 5’ – 3’ (green lines) or 3’ – 5’ (red lines) orientation. Gaps in the mapping of individual reads (dotted lines) indicate the putative position of introns. (iii) Coverage of mapped reads (y-axis, in reverse) also indicates the position of putative exons. (iv) RT-PCR revealed that ETH_00017050 and ETH_00017055 are actually part of a single ethap2 transcript. Exons of this transcript are indicated as blue boxes, while separating introns are indicated as a blue line. The resulting EtHAP2 coding sequence reads from right (5’) to left (3’). (b) Multiple alignment of amino acid sequences corresponding to the HAP2 domains of E. tenella (EtHAP2), T. gondii (TGME49_285940, TgHAP2), P. berghei (PBANKA_121260, PbHAP2) and C. reinhardtii (C_530033, CrHAP2). Blue shading is proportional to levels of conservation between the four species. Amino acid sequence limits are indicated to the right. (c) The domain architecture of the EtHAP2 protein is illustrated (to scale). An N-terminal signal peptide is followed by an extracellular region (comprising the HAP2 domain), a transmembrane domain (TM), and a C-terminal intracellular domain containing a lysine/arginine-rich domain (KR).