Deletion of a novel protein kinase with PX and FYVE‐related domains increases the rate of differentiation of Trypanosoma brucei

Growth control of African trypanosomes in the mammalian host is coupled to differentiation of a non‐dividing life cycle stage, the stumpy bloodstream form. We show that a protein kinase with novel domain architecture is important for growth regulation. Zinc finger kinase (ZFK) has a kinase domain related to RAC and S6 kinases flanked by a FYVE‐related zinc finger and a phox (PX) homology domain. To investigate the function of the kinase during cyclical development, a stable transformation procedure for bloodstream forms of differentiation‐competent (pleomorphic) Trypanosoma brucei strains was established. Deletion of both allelic copies of ZFK by homologous recombination resulted in reduced growth of bloodstream‐form parasites in culture, which was correlated with an increased rate of differentiation to the non‐dividing stumpy form. Growth and differentiation rates were returned to wild‐type level by ectopic ZFK expression. The phenotype is stage‐specific, as growth of procyclic (insect form) trypanosomes was unaffected, and Δzfk/Δzfk clones were able to undergo full cyclical development in the tsetse fly vector. Deletion of ZFK in a differentiation‐defective (monomorphic) strain of T. brucei did not change its growth rate in the bloodstream stage. This suggests a function of ZFK associated with the trypanosomes' decision between either cell cycle progression, as slender bloodstream form, or differentiation to the non‐dividing stumpy form.

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