Trypanosoma brucei, the parasite responsible for causing African Sleeping Sickness, has many unusual biochemical features and processes, including RNA Polymerase II (RNAPII)-dependent gene transcription. We have identified a trypanosome-specific protein that is associated with the RNAPII transcriptional machinery, is found in the nucleus, and is essential for parasite growth as assessed by RNA interference.  This protein, named Essential Nuclear Factor (ENF), contains motifs associated with known transcription factors, but the protein is unique to trypanosomes. Using biotinylated DNA probes and trypanosome nuclear extract, which contains Trypanosoma brucei ENF (TbENF), we have found that TbENF49 interacts with DNA. To test the involvement of TbENF in transcription, RNA levels will be compared between TbENF49 RNAi-induced  and non-induced cell lines.  Furthermore, a TAP-tagged TbENF49 cell line has been generated and results of protein interaction studies will be presented.

Characterization of an essential trypanosome DNA binding protein

244th National Meeting (2012)

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