A complex small rna repertoire is generated by a plant/fungal-like machinery and effected by a metazoan-like argonaute in the single-cell human parasite toxoplasma gondii

In rna silencing,small rnas produced by the rnase-iii dicer guide argonaute-like proteins as part of rna-induced silencing complexes (risc) to regulate gene expression transcriptionally or post transcriptionally. here,we have characterized the rna silencing machinery and exhaustive small rnaome of toxoplasma gondii,member of the apicomplexa,a phylum of animal- and human-infecting parasites that cause extensive health and economic damages to human populations worldwide. remarkably,the small rna-generating machinery of toxoplasma is phylogenetically and functionally related to that of plants and fungi,and accounts for an exceptionally diverse array of small rnas. this array includes conspicuous populations of repeat-associated small interfering rna (sirna),which,as in plants,likely generate and maintain heterochromatin at dna repeats and satellites. toxoplasma small rnas also include many micrornas with clear metazoanlike features whose accumulation is sometimes extremely high and dynamic,an unexpected finding given that toxoplasma is a unicellular protist. both plant-like heterochromatic small rnas and metazoan-like micrornas bind to a single argonaute protein,tg-ago. toxoplasma mirnas co-sediment with polyribosomes,and thus,are likely to act as translational regulators,consistent with the lack of catalytic residues in tg-ago. mass spectrometric analyses of the tg-ago protein complex revealed a common set of virtually all known risc components so far characterized in human and drosophila,as well as novel proteins involved in rna metabolism. in agreement with its loading with heterochromatic small rnas,tg-ago also associates substoichiometrically with components of known chromatin-repressing complexes. thus,a puzzling patchwork of silencing processor and effector proteins from plant,fungal and metazoan origin accounts for the production and action of an unsuspected variety of small rnas in the single-cell parasite toxoplasma and possibly in other apicomplexans. this study establishes toxoplasma as a unique model system for studying the evolution and molecular mechanisms of rna silencing among eukaryotes. © 2010 braun et al.