TFIIE orchestrates the recruitment of the TFIIH kinase module at promoter before release during transcription.

In eukaryotes, the general transcription factors TFIIE and TFIIH assemble at the transcription start site with RNA Polymerase II. However, the mechanism by which these transcription factors incorporate the preinitiation complex and coordinate their action during RNA polymerase II transcription remains elusive. Here we show that the TFIIEα and TFIIEß subunits anchor the TFIIH kinase module (CAK) within the preinitiation complex. In addition, we show that while RNA polymerase II phosphorylation and DNA opening occur, CAK and TFIIEα are released from the promoter. This dissociation is impeded by either ATP-γS or CDK7 inhibitor THZ1, but still occurs when XPB activity is abrogated. Finally, we show that the Core-TFIIH and TFIIEß are subsequently removed, while elongation factors such as DSIF are recruited. Remarkably, these early transcriptional events are affected by TFIIE and TFIIH mutations associated with the developmental disorder, trichothiodystrophy.

Leishmania infantum EndoG is an endo/exo-nuclease essential for parasite survival.

EndoG, a member of the DNA/RNA non-specific ßßα-metal family of nucleases, has been demonstrated to be present in many organisms, including Trypanosomatids. This nuclease participates in the apoptotic program in these parasites by migrating from the mitochondrion to the nucleus, where it takes part in the degradation of genomic DNA that characterizes this process. We now demonstrate that Leishmania infantum EndoG (LiEndoG) is an endo-exonuclease that has a preferential 5' exonuclease activity on linear DNA. Regardless of its role during apoptotic cell death, this enzyme seems to be necessary during normal development of the parasites as indicated by the reduced growth rates observed in LiEndoG hemi-knockouts and their poor infectivity in differentiated THP-1 cells. The pro-life role of this protein is also corroborated by the higher survival rates of parasites that over-express this protein after treatment with the LiEndoG inhibitor Lei49. Taken together, our results demonstrate that this enzyme plays essential roles in both survival and death of Leishmania parasites.

5'-Trityl-substituted thymidine derivatives as a novel class of antileishmanial agents: Leishmania infantum EndoG as a potential target.

Two series of 5'-triphenylmethyl (trityl)-substituted thymidine derivatives were synthesized and tested against Leishmania infantum axenic promastigotes and amastigotes. Several of these compounds show significant antileishmanial activity, with IC50 values in the low micromolar range. Among these, 3'-O-(isoleucylisoleucyl)-5'-O-(3,3,3-triphenylpropanoyl)thymidine displays particularly good activity against intracellular amastigotes. Assays performed to characterize the nature of parasite cell death in the presence of the tritylthymidines indicated significant alterations in mitochondrial transmembrane potential, an increase in superoxide concentrations, and also significant decreases in DNA degradation during the cell death process. Results point to the mitochondrial nuclease LiEndoG as a target for the action of this family of compounds.

Mitochondrial dysfunction in Trypanosoma cruzi: the role of Serratia marcescens prodigiosin in the alternative treatment of Chagas disease.

BACKGROUND: Chagas disease is a health threat for many people, mostly those living in Latin America. One of the most important problems in treatment is the limitation of existing drugs. Prodigiosin, produced by Serratia marcescens (Rhodnius prolixus endosymbiont), belongs to the red-pigmented bacterial prodiginine family, which displays numerous biological activities, including antibacterial, antifungal, antiprotozoal, antimalarial, immunosuppressive, and anticancer properties. Here we describe its effects on Trypanosoma cruzi mitochondria belonging to Tc I and Tc II. RESULTS: Parasites exposed to prodigiosin altered the mitochondrial function and oxidative phosphorylation could not have a normal course, probably by inhibition of complex III. Prodigiosin did not produce cytotoxic effects in lymphocytes and Vero cells and has better effects than benznidazole. Our data suggest that the action of prodigiosin on the parasites is mediated by mitochondrial structural and functional disruptions that could lead the parasites to an apoptotic-like cell death process. CONCLUSIONS: Here, we propose a potentially useful trypanocidal agent derived from knowledge of an important aspect of the natural life cycle of the parasite: the vector-parasite interaction. Our results indicate that prodigiosin could be a good candidate for the treatment of Chagas disease.

[Myocardial cellular damage and the activity of the mitochondrial ATP synthase in rats infected with a Colombian strain of Trypanosoma cruzi]. / Lesión celular del miocardio y actividad de la ATPsintasa mitocondrial en ratas infectadas con una cepa colombiana de Trypanosoma cruzi.

INTRODUCTION: Chagas disease is the main cause of cardiomyopathy in endemic regions of Latin America. Alterations in the cardiac mitochondrial energy metabolism caused by Trypanosoma cruzi can be involved in the development of this cardiomyopathy during the course of Chagas disease. OBJECTIVE: The cellular injury of the rat myocardium was investigated in rats infected with the Colombian Mg8 strain of Trypanosoma cruzi. The activity of mitochondrial ATP synthase was measured to determine the relationship heart damage with the energy metabolism. MATERIALS AND METHODS: Two groups of five rats each were infected with tripomastigotes, with 1 group of 6 rats serving as controls. The course of infection was characterized by parasitological, histopathological and molecular studies. The mitochondrial ATP synthase activity of the myocardium was evaluated in all rats. RESULTS: Peak parasitaemia (day 26 post infection) and the time of parasite clearance from circulating blood (day 60 post infection) were determined for acute and chronic phase models. The histopathological and molecular results showed that the Colombian Mg8 strain has tropism to the cardiac tissue and causes considerable cellular injury of the myocardium in rats during both phases. Despite the lesions observed in infected rats, no statistical difference in the activity of the mitochondrial ATPsynthase was observed between them and the non-infected rats. CONCLUSION: Mitochondrial energy metabolism of the cardiomyocites does not appear to change during cellular injury of rat myocardium associated with infection by the Colombian Mg8 T. cruzi strain.