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Leishmania  and  Trypanosoma  are unicellular parasites that possess markedly original biological features as compared to other eukaryotes. The  Leishmania  genome displays a constitutive ‘mosaic aneuploidy’, whereas in  Trypanosoma brucei , the megabase-sized chromosomes are diploid. We accurately analysed DNA replication parameters in three  Leishmania  species and  Trypanosoma brucei  as well as mouse embryonic fibroblasts (MEF). Active replication origins were visualized at the single molecule level using DNA molecular combing. More than one active origin was found on most DNA fibres, showing that the chromosomes are replicated from multiple origins. Inter-origin distances (IODs) were measured and found very large in trypanosomatids: the mean IOD was 160 kb in  T. brucei  and 226 kb in  L. mexicana . Moreover, the progression of replication forks was faster than in any other eukaryote analyzed so far (mean velocity 1.9 kb/min in  T. brucei  and 2.4–2.6 kb/min in  Leishmania ). The estimated total number of active DNA replication origins in trypanosomatids is ~170. Finally, 14.4% of unidirectional replication forks were observed in  T. brucei , in contrast to 1.5–1.7% in  Leishmania  and 4% in MEF cells. The biological significance of these original features is discussed.

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Single-molecule analysis of DNA replication reveals novel features in the divergent eukaryotes Leishmania and Trypanosoma brucei versus mammalian cells