A Genome-Wide Tethering Screen Reveals Novel Potential Post-Transcriptional Regulators in Trypanosoma brucei
Figure 3
(A) Sequence reads that were linked to the lambda-N peptide were extracted, mapped onto the trypanosome genome and visualised using the Integrative Genomics Viewer (IGV) tool. The Figure shows the sequence coverage plots for a ∼12 kbp region of chromosome 5. Open reading frames are in purple and mRNA untranslated regions in dark grey. Reads were derived from tetracycline-induced cells grown without blasticidin (upper panel), or with the 10x blasticidin concentration (bottom panel). For better visualization, the plotted Y scales display different range values. In the 10x blasticidin plot, note the cluster over ZC3H11; the isolated “pile” downstream of Tb927.5.820 would be removed later in the filtering process. (B) These are the same results as in (A), but in close-up focusing on the ZC3H11 (Tb927.5.810) open reading frame. Each “stack” represents a unique target fragment, mapped to forward (plus) and reverse (minus) strands, for induced cells grown without (upper panel) and with (bottom panel) blasticidin pressure. Light blue boxes indicate the ZC3H11 coding sequence (transcribed left to right) while zinc finger domain and HNPY motif (residues 186–189) are indicated in green and orange respectively. The minimum region that was required for activity in tethering assays was the C-terminal part, starting at amino acid 186.