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A more recent version of this article appeared on October 1, 2002
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Submitted on May 8, 2002
Revised on July 11, 2002
Accepted on July 22, 2002
1 School of Biological Sciences, University of Manchester, Oxford Road, Manchester M13 9PT, UK
* Corresponding author. E-mail address: keith.matthews{at}man.ac.uk.
Life cycle-differentiation of African trypanosomes entails developmental regulation of mitochondrial activity. This requires regulation of the nuclear genome and the kinetoplast, the trypanosome's unusual mitochondrial genome. To investigate the potential cross-talk between the nuclear and mitochondrial genome during the events of differentiation we have (i) disrupted expression of a nuclear-encoded component of the cytochrome oxidase (COX) complex and (ii) generated dyskinetoplastid cells, which lack a mitochondrial genome. Using RNA interference (RNAi) and by disrupting the nuclear COX VI gene we demonstrate independent regulation of COX component mRNAs encoded in the nucleus and kinetoplast. However, two independent approaches (acriflavine treatment and RNAi ablation of mitochondrial topoisomerase II) failed to establish clonal lines of dyskinetoplastid bloodstream forms. Nevertheless, dyskinetoplastid forms generated in vivo could undergo two life cycle differentiation events: transition from bloodstream slender to stumpy forms and the initiation of transformation to procyclic forms. However, they subsequently arrested at a specific point in this developmental programme prior to cell cycle re-entry. These results provide strong evidence for a requirement for kDNA in the bloodstream and for a kinetoplast-dependent control point during differentiation to procyclic forms.
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