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Natalie M McAdams, Rachel M Simpson, Runpu Chen, Yijun Sun, Laurie Read
The trypanosome RNA Editing Substrate Binding Complex (RESC) acts as the platform for mitochondrial uridine insertion/deletion RNA editing and facilitates the protein-protein and protein-RNA interactions required for the editing process. RESC is broadly comprised of two subcomplexes: GRBC (Guide RNA Binding Complex) and REMC (RNA Editing Mediator Complex). Here, we characterize the function and position in RESC organization of a previously unstudied RESC protein, MRB7260. We show that MRB7260 forms numerous RESC-related complexes, including a novel, small complex with the guide RNA binding protein, GAP1, which is a canonical GRBC component, and REMC components MRB8170 and TbRGG2...
January 12, 2018: RNA
Qunbo Tong, Rui Chen, Qingming Kong, Julie Goossens, Magdalena Radwanska, Di Lou, Jianzu Ding, Bin Zheng, Yixiu Fu, Tianping Wang, Magez Stefan, Shaohong Lu
Trypanosoma evansi (T. evansi) is the most widely spread pathogenic trypanosome in the world. The control of trypanosomiasis depends on accurate diagnosis and effective treatment. Focusing on the presence of T. evansi in Asia, we developed a detection assay based on tracing phosphate ions (Pi) generated during LAMP targeting the variant surface glycoprotein (VSG) gene of Rode Trypanozoon antigenic type 1.2 (RoTat 1.2 VSG). The diagnostic potential as well as the use of the assay as a test-of-cure method after berenil treatment, was assessed in mice at different time points of infection...
January 30, 2018: Veterinary Parasitology
David Izuchukwu Ugwu, Uchechukwu Chris Okoro, Narendra Kumar Mishra
The reported toxicities of current antitrypanosomal drugs and the emergence of drug resistant trypanosomes underscore the need for the development of new antitrypanosomal agents. We report herein the synthesis and antitrypanosomal activity of 24 new amide derivatives of 3-aminoquinoline, bearing substituted benzenesulphonamide. Nine of the new derivatives showed comparable antitrypanosomal activities at IC50 range of 1-6 nM (melarsoprol 5 nM). Compound 11n and 11v are more promising antitrypanosomal agents with IC50 1...
2018: PloS One
Israel Canela-Pérez, Imelda López-Villaseñor, Ana María Cevallos, Roberto Hernández
Trypanosoma cruzi is the aetiologic agent of Chagas disease. Our research group studies ribosomal RNA (rRNA) gene transcription and nucleolus dynamics in this species of trypanosomes. RPA31 is an essential subunit of RNA polymerase I (Pol I) whose presence is apparently restricted to trypanosomes. Using fluorescent-tagged versions of this protein (TcRPA31-EGFP), we describe its nuclear distribution during growth and metacyclogenesis. Our findings indicate that TcRPA31-EGFP alters its nuclear presence from concentrated nucleolar localization in exponentially growing epimastigotes to a dispersed granular distribution in the nucleoplasm of stationary epimastigotes and metacyclic trypomastigotes...
January 10, 2018: Parasitology Research
Julianne Mendi Muthinja, Johanna Ripp, Timothy Krüger, Andrea Imle, Tamás Haraszti, Oliver T Fackler, Joachim P Spatz, Markus Engstler, Friedrich Frischknecht
Motile cells and pathogens migrate in complex environments and yet are mostly studied on simple 2D substrates. In order to mimic the diverse environments of motile cells a set of assays including substrates of defined elasticity, microfluidics, micropatterns, organotypic cultures and 3D gels have been developed. We briefly introduce these and then focus on the use of micro-patterned pillar arrays, which help to bridge the gap between 2D and 3D. These structures are made from polydimethylsiloxane, a moldable plastic and their use has revealed new insights into mechanoperception in Caenorhabditis elegans, gliding motility of Plasmodium, swimming of trypanosomes and nuclear stability in cancer cells...
January 7, 2018: Cellular Microbiology
Lynne M Harris, Katelyn R Monsell, Florian Noulin, M Toyin Famodimu, Nicolas Smargiasso, Christian Damblon, Paul Horrocks, Catherine J Merrick
G-quadruplexes are DNA or RNA secondary structures that can be formed from guanine-rich nucleic acids. These four-stranded structures, composed of stacked quartets of guanine bases, can be highly stable and have been demonstrated to occur in vivo in the DNA of human cells and other systems, where they play important biological roles, influencing processes such as telomere maintenance, DNA replication and transcription, or in the case of RNA G-quadruplexes, RNA translation and processing. We report for the first time that DNA G-quadruplexes can be detected in the nuclei of the malaria parasite Plasmodium falciparum, which has one of the most A/T-biased genomes sequenced and therefore possesses few guanine-rich sequences with the potential to form G-quadruplexes...
January 8, 2018: Antimicrobial Agents and Chemotherapy
Paula Boeira Bassi, Fernanda Fortes de Araújo, Guilherme Caetano Garcia, Marcos Vinícius da Silva, Carlo José Freire Oliveira, Eustáquio Resende Bittar, Matheus de Souza Gomes, Laurence Rodrigues do Amaral, Matheus Fernandes Costa E Silva, Gabriel Antônio Nogueira Nascentes, Virmondes Rodrigues Junior, Olindo Assis Martins-Filho, Márcio Sobreira Silva Araújo, Joely Ferreira Figueiredo Bittar
Trypanosoma vivax infection causes relevant economical impact due to high morbidity and mortality leading to negative impact on local livestock. Despite parasitological and serological methods are used for the diagnosis of T. vivax infection, gaps regarding sensitivity and specificity of these methods still represent a challenge. The present study aimed to compare the kinetics of parasitological and serological parameters in cattle experimentally infected with T. vivax along with immunophenotypic analysis of whole blood leukocytes...
January 5, 2018: Experimental Parasitology
José Bubis, Juan Carlos Martínez, Maritza Calabokis, Joilyneth Ferreira, Carlos E Sanz-Rodríguez, Victoria Navas, José Leonardo Escalona, Yurong Guo, Susan S Taylor
The full gene sequence encoding for the Trypanosoma equiperdum ortholog of the cAMP-dependent protein kinase (PKA) regulatory (R) subunits was cloned. A poly-His tagged construct was generated [TeqR-like(His)8], and the protein was expressed in bacteria and purified to homogeneity. The size of the purified TeqR-like(His)8 was determined to be ∼ 57,000 Da by molecular exclusion chromatography indicating that the parasite protein is a monomer. Limited proteolysis with various proteases showed that the T. equiperdum R-like protein possesses a hinge region very susceptible to proteolysis...
December 27, 2017: Biochimie
Sen Claudine Henriette Ngomtcho, Judith Sophie Weber, Elisabeth Ngo Bum, Thaddeus Terlumun Gbem, Sørge Kelm, Mbunkah Daniel Achukwi
BACKGROUND: African trypanosomes are mainly transmitted through the bite of tsetse flies (Glossina spp.). The present study investigated the occurrence of pathogenic trypanosomes in tsetse flies and cattle in tsetse fly-infested areas of Northern Cameroon. RESULTS: Trypanosomes were identified using nested polymerase chain reaction (PCR) analysis of internal transcribed spacer 1 (ITS1) region, both by size estimation and sequencing of PCR products. Apparent density indices recorded in Gamba and Dodeo were 3...
December 29, 2017: Parasites & Vectors
Sandro Käser, Mathilde Willemin, Felix Schnarwiler, Bernd Schimanski, Daniel Poveda-Huertes, Silke Oeljeklaus, Beat Haenni, Benoît Zuber, Bettina Warscheid, Chris Meisinger, André Schneider
Mitochondria cannot form de novo but require mechanisms that mediate their inheritance to daughter cells. The parasitic protozoan Trypanosoma brucei has a single mitochondrion with a single-unit genome that is physically connected across the two mitochondrial membranes with the basal body of the flagellum. This connection, termed the tripartite attachment complex (TAC), is essential for the segregation of the replicated mitochondrial genomes prior to cytokinesis. Here we identify a protein complex consisting of three integral mitochondrial outer membrane proteins-TAC60, TAC42 and TAC40-which are essential subunits of the TAC...
December 2017: PLoS Pathogens
Diana Ortiz, W Armand Guiguemde, Jared T Hammill, Angela K Carrillo, Yizhe Chen, Michele Connelly, Kayla Stalheim, Carolyn Elya, Alex Johnson, Jaeki Min, Anang Shelat, David C Smithson, Lei Yang, Fangyi Zhu, R Kiplin Guy, Scott M Landfear
Leishmaniasis is a parasitic infection that afflicts approximately 12 million people worldwide. There are several limitations to the approved drug therapies for leishmaniasis, including moderate to severe toxicity, growing drug resistance, and the need for extended dosing. Moreover, miltefosine is currently the only orally available drug therapy for this infection. We addressed the pressing need for new therapies by pursuing a two-step phenotypic screen to discover novel, potent, and orally bioavailable antileishmanials...
December 29, 2017: PLoS Neglected Tropical Diseases
Noelia Lander, Miguel A Chiurillo, Mayara S Bertolini, Roberto Docampo, Aníbal E Vercesi
The presence of a conserved mechanism for mitochondrial calcium uptake in trypanosomatids was crucial for the molecular identification of the mitochondrial calcium uniporter (MCU), a long-sought channel present in most eukaryotic organisms. Since then, research efforts to elucidate the role of MCU and its regulatory elements in different biological models have multiplied. MCU is the pore-forming subunit of a multimeric complex (the MCU complex or MCUC) and its predicted structure in trypanosomes is simpler than in mammalian cells, lacking two of its subunits and probably possessing other unidentified components...
December 29, 2017: Cell Biology International
Felipe Gazos-Lopes, Jessica L Martin, Peter C Dumoulin, Barbara A Burleigh
Intracellular infection and multi-organ colonization by the protozoan parasite, Trypanosoma cruzi, underlie the complex etiology of human Chagas disease. While T. cruzi can establish cytosolic residence in a broad range of mammalian cell types, the molecular mechanisms governing this process remain poorly understood. Despite the anticipated capacity for fatty acid synthesis in this parasite, recent observations suggest that intracellular T. cruzi amastigotes may rely on host fatty acid metabolism to support infection...
December 27, 2017: PLoS Pathogens
Jeiczon Jaimes-Dueñez, Omar Triana-Chávez, Ana M Mejía-Jaramillo
Animal Trypanosomiasis (AT) is one of the most important problems in the Colombian livestock industry reducing its production around 30%. Caribbean and Orinoquia regions play a significant role in the development of this industry, having about 6.9 million cattle and 113,000 buffaloes. Considering the paucity in studies to understand the epidemiological features and control of AT in Colombia, the present study reports the seasonal transmission patterns and phylogeographic traits of the causal agents of AT in cattle and buffaloes from these regions...
January 15, 2018: Veterinary Parasitology
S Andrea Moreno, Gabriela V Cantos
We hypothesize that the hexokinases of trypanosomes of the subgenus Trypanozoon match the blood glucose levels of hosts. We studied the kinetic properties of purified hexokinase in T. equiperdum (specific activity=302U/mg), and compare with other members of Trypanozoon. With ATP (Km=104.7μM) as phosphate donor, hexokinase catalyzes the phosphorylation of glucose (Km=24.9μM) and mannose (Km=8.8μM). With respect to glucose, mannose and inorganic pyrophosphate respectively are a competitive, and a mixed inhibitor of hexokinase...
December 20, 2017: Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology
Whitney Bullard, Rudo Kieft, Robert Sabatini
Recently, 5-hydroxymethyluracil (5hmU) was identified in mammalian genomic DNA as an oxidative product of thymine by the ten-eleven translocation (TET) proteins. While the biological role of this modification remains unclear, identifying its genomic location will assist in elucidating function. Here we present a rapid and robust method to selectively tag and enrich genomic regions containing 5hmU. This method involves the selective glucosylation of 5hmU residues by the base J glucosyltransferase from trypanosomes creating glucosylhydroxymethyluracil (base J)...
January 2017: Biology Methods and Protocols
Pythagore Soubgwi Fogue, Flobert Njiokou, Gustave Simo
Despite the economic impact of trypanosome infections, few investigations have been undertaken on the population genetics and transmission dynamics of animal trypanosomes. In this study, microsatellite markers were used to investigate the population genetics of Trypanosoma congolense "forest type", with the ultimate goal of understanding its transmission dynamics between tsetse flies and domestic animals. Blood samples were collected from pigs, sheep, goats and dogs in five villages in Fontem, South-West region of Cameroon...
2017: Parasite: Journal de la Société Française de Parasitologie
Rosemary Bateta, Jingwen Wang, Yineng Wu, Brian L Weiss, Wesley C Warren, Grace A Murilla, Serap Aksoy, Paul O Mireji
BACKGROUND: Tsetse flies (Glossina spp.) are the prominent vector of African trypanosome parasites (Trypanosoma spp.) in sub-Saharan Africa, and Glossina pallidipes is the most widely distributed species in Kenya. This species displays strong resistance to infection by parasites, which are typically eliminated in the midgut shortly after acquisition from the mammalian host. Although extensive molecular information on immunity for the related species Glossina morsitans morsitans exists, similar information is scarce for G...
December 19, 2017: Parasites & Vectors
Sylvain Trépout, Anne-Marie Tassin, Sergio Marco, Philippe Bastin
The protist Trypanosoma brucei is an emerging model for the study of cilia and flagella. Here, we used scanning transmission electron microscopy (STEM) tomography to describe the structure of the trypanosome transition zone (TZ). At the base of the TZ, nine transition fibres irradiate from the B microtubule of each doublet towards the membrane. The TZ adopts a 9+0 structure throughout its length of ∼300 nm and its lumen contains an electron-dense structure. The proximal portion of the TZ has an invariant length of 150 nm and is characterised by a collarette surrounding the membrane and the presence of electron-dense material between the membrane and the doublets...
December 14, 2017: Journal of Structural Biology
Xuan Zhang, Xiao-Kun Hong, Su-Jin Li, De-Hua Lai, Geoff Hide, Zhao-Rong Lun, Yan-Zi Wen
Trypanosoma musculi, a common blood flagellate found in mice, is similar in morphology and life cycle to the rat trypanosome T. lewisi. Both species belong to the subgenus Herpetosoma, and as T. lewisi has recently been shown to be a zoonotic pathogen, there is concern that T. musculi could also be potentially infective to humans. To test this hypothesis, a well-established method, the normal human serum (NHS) incubation test, was carried out which distinguishes human and non-human infective trypanosomes...
December 13, 2017: Experimental Parasitology
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