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Molecular and Biochemical Parasitology

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https://www.readbyqxmd.com/read/28522152/anti-trypanosomal-activity-of-cationic-n-heterocyclic-carbene-gold-i-complexes
#1
Isabel Winter, Julia Lockhauserbäumer, Gertrud Lallinger-Kube, Rainer Schobert, Klaus Ersfeld, Bernhard Biersack
Two gold(I) N-heterocyclic carbene complexes 1a and 1b were tested for their anti-trypanosomal activity against Trypanosoma brucei parasites. Both gold compounds exhibited excellent anti-trypanosomal activity (IC50=0.9-3.0nM). The effects of the gold complexes 1a and 1b on the T. b. brucei cytoskeleton were evaluated. Rapid detachment of the flagellum from the cell body occurred after treatment with the gold complexes. In addition, a quick and complete degeneration of the parasitic cytoskeleton was induced by the gold complexes, only the microtubules of the detached flagellum remained intact...
May 15, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28438458/an-efficient-cumate-inducible-system-for-procyclic-and-bloodstream-form-trypanosoma-brucei
#2
Feng-Jun Li, Zhi-Shen Xu, Htay Mon Aye, Anaïs Brasseur, Zhao-Rong Lun, Kevin S W Tan, Cynthia Y He
In Trypanosoma brucei, the tetracycline-inducible system enables tightly-regulated, highly-efficient expression of recombinant proteins or double-stranded RNA in both procyclic and bloodstream form cells, providing useful molecular genetic tools to study gene functions. An alternative, vanillic acid-inducible system is recently described for procyclic T. brucei, providing ∼18-fold increase in GFP reporter expression upon induction (Sunter JD. Mol. Biochem. Parasitol. 2016, 207:45-48). Here we describe a cumate-inducible system that allows efficient, tunable gene expression showing >300-fold increase in GFP expression upon induction...
April 21, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28427949/parasitism-finds-many-solutions-to-the-same-problems-in-red-algae-florideophyceae-rhodophyta
#3
Jillian M Freese, Christopher E Lane
Parasitic red algae evolve from a common ancestor with their hosts, parasitizing cousins using familiar cellular mechanisms. They have independently evolved over one hundred times within the exclusively multicellular red algal class Florideophyceae. Reduced morphology, a lack of pigmentation, and direct cell-cell connections with their hosts are markers of red algal parasitism. With so many potential evolutionary pathways, red algal parasite diversity offers a unique test case to understand the earliest stages of this lifestyle transition...
April 17, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28414172/lccl-protein-complex-formation-in-plasmodium-is-critically-dependent-on-lap1
#4
Annie Z Tremp, Vikram Sharma, Victoria Carter, Edwin Lasonder, Johannes T Dessens
Successful sporogony of Plasmodium berghei in vector mosquitoes requires expression of a family of six modular proteins named LCCL lectin domain adhesive-like proteins (LAPs). The LAPs share a subcellular localization in the crystalloid, a unique parasite organelle that forms during ookinete development. Here, LAP interactions in P. berghei were studied using a series of parasite lines stably expressing reporter-tagged LAPs combined with affinity purification and high accuracy label free quantitative mass spectrometry...
April 13, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28392476/schistosoma-mansoni-displays-an-adenine-phosphoribosyltransferase-preferentially-expressed-in-mature-female-gonads-and-vitelaria
#5
Ana Eliza Zeraik, Vitor Hugo Balasco Serrão, Larissa Romanello, Juliana Roberta Torini, Alexandre Cassago, Ricardo DeMarco, Humberto D'Muniz Pereira
Schistosoma mansoni depends upon the purine salvage pathway to obtain purine nucleotides; therefore, enzymes from this pathway are essential for parasite survival. Here, we focused on the adenine phosphoribosyltransferase (APRT) enzyme, which catalyzes the condensation reaction between adenine and PRPP (5-phosphoribosylpyrophosphate) to produce AMP and PPi. Kinetic experiments using the heterologously expressed protein of one APRT isoform from S. mansoni indicate that it is catalytically active, and whole-mount in situ hybridization studies indicate that the transcripts of this protein are concentrated in the posterior region of the ovary and vitellaria of female adult worms...
April 6, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28389272/characterization-of-ciliobrevin-a-mediated-dynein-atpase-inhibition-on-flagellar-motility-of-leishmania-donovani
#6
G Srinivas Reddy, Aakash Gautam Mukhopadhyay, Chinmoy Sankar Dey
Axonemal dyneins are members of AAA+ proteins involved in force generation and are responsible for flagellar motility in eukaryotes. In this study, we characterized the effects of ciliobrevin A (CbA), a dynein ATPase inhibitor, on flagella driven motility of the protozoan parasite Leishmania donovani. Using fast-capture video microscopy, we observed that CbA decreased flagellar beat frequency of swimming parasites in a concentration-dependent manner. Beat frequency of live and reactivated L. donovani decreased by approximately 89% and 41% respectively in the presence of 250μM CbA...
April 5, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28385564/genome-wide-identification-of-microrna-targets-in-the-neglected-disease-pathogens-of-the-genus-echinococcus
#7
Natalia Macchiaroli, Lucas L Maldonado, Magdalena Zarowiecki, Marcela Cucher, María Inés Gismondi, Laura Kamenetzky, Mara Cecilia Rosenzvit
MicroRNAs (miRNAs), a class of small non-coding RNAs, are key regulators of gene expression at post-transcriptional level and play essential roles in biological processes such as development. MiRNAs silence target mRNAs by binding to complementary sequences in the 3'untranslated regions (3'UTRs). The parasitic helminths of the genus Echinococcus are the causative agents of echinococcosis, a zoonotic neglected disease. In previous work, we performed a comprehensive identification and characterization of Echinococcus miRNAs...
April 3, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28385563/evidence-of-putative-non-coding-rnas-from-leishmania-untranslated-regions
#8
Felipe Freitas Castro, Patricia C Ruy, Karina Nogueira Zeviani, Ramon Freitas Santos, Juliano Simões Toledo, Angela Kaysel Cruz
Non-coding RNAs (ncRNAs) are regulatory elements present in a wide range of organisms, including trypanosomatids. ncRNAs transcribed from the untranslated regions (UTRs) of coding genes have been described in the transcriptomes of several eukaryotes, including Trypanosoma brucei. To uncover novel putative ncRNAs in two Leishmania species, we examined a L. major cDNA library and a L. donovani non-polysomal RNA library. Using a combination of computational analysis and experimental approaches, we classified 26 putative ncRNA in L...
April 3, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28373094/inducible-protein-stabilization-system-in-leishmania-mexicana
#9
Lucie Podešvová, Huan Huang, Vyacheslav Yurchenko
Targeted regulation of protein levels is an important tool to investigate the role of proteins essential for cell function and development. In recent years, methods based on the Escherichia coli dihydrofolate reductase destabilization domain (ecDHFR DD) have been established and used in various cell types. ecDHFR DD destabilizes the fused protein of interest and causes its degradation by proteasomes, unless it is stabilized by a specific ligand, trimethoprim. In this work we developed an inducible protein stabilization system in Leishmania mexicana based on ecDHFR DD...
March 31, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28373093/experimental-and-bioinformatic-characterization-of-cabp2933-an-ef-hand-protein-of-giardia-intestinalis
#10
Magda E Alvarado, Claudia Rubiano, Eliana Calvo, Vanessa Gómez, Moisés Wasserman
Giardia intestinalis is a parasite that inhabits the small intestine of humans. This parasite is a divergent eukaryote with a compact genome. The calcium ion is an essential messenger in cell signaling. Calcium's role as a messenger is mediated through calcium-binding proteins (CaBPs) that decode the message. The most important family of CaBPs is the EF-Hand protein family. In this study we have explored the role of EF-Hand protein CaBP2933. We analyzed its location, confirmed its ability to bind calcium and identified some of its interacting proteins...
March 31, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28366668/the-intermembrane-space-protein-erv1-of-trypanosoma-brucei-is-essential-for-mitochondrial-fe-s-cluster-assembly-and-operates-alone
#11
Alexander C Haindrich, Michala Boudová, Marie Vancová, Priscila Peña Diaz, Eva Horáková, Julius Lukeš
Sulfhydryl oxidase Erv1 is a ubiquitous and conserved protein of the mitochondrial intermembrane space that plays a role in the transport of small sulfur-containing proteins. In higher eukaryotes, Erv1 interacts with the mitochondrial import protein Mia40. However, Trypanosoma brucei lacks an obvious Mia40 homologue in its genome. Here we show by tandem affinity purification and mass spectrometry that in this excavate protist, Erv1 functions without a Mia40 homologue and most likely any other interaction partner...
March 31, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28366667/characterization-and-role-of-the-3-methylglutaconyl-coenzyme-a-hidratase-in-trypanosoma-brucei
#12
Mariana Leão de Lima Stein, Marcelo Yudi Icimoto, Erica Valadares de Castro Levatti, Vitor Oliveira, Anita Hilda Straus, Sergio Schenkman
Trypanosoma brucei, the agent of African Trypanosomiasis, is a flagellated protozoan parasite that develops in tsetse flies and in the blood of various mammals. The parasite acquires nutrients such as sugars, lipids and amino acids from their hosts. Amino acids are used to generate energy and for protein and lipid synthesis. However, it is still unknown how T. brucei catabolizes most of the acquired amino acids. Here we explored the role of an enzyme of the leucine catabolism, the 3-methylglutaconyl-Coenzyme A hydratase (3-MGCoA-H)...
March 31, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28356223/the-role-of-the-pi-3-5-p2-kinase-tbfab1-in-endo-lysosomal-trafficking-in-trypanosoma-brucei
#13
Julia K Gilden, Khan Umaer, Emilia K Kruzel, Oliver Hecht, Renan O Correa, John M Mansfield, James D Bangs
Protein trafficking through endo/lysosomal compartments is critically important to the biology of the protozoan parasite Trypanosoma brucei, but the routes material may take to the lysosome, as well as the molecular factors regulating those routes, remain incompletely understood. Phosphoinositides are signaling phospholipids that regulate many trafficking events by recruiting specific effector proteins to discrete membrane subdomains. In this study, we investigate the role of one phosphoinositide, PI(3,5)P2 in T...
March 27, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28347672/structural-and-kinetic-analysis-of-schistosoma-mansoni-adenylosuccinate-lyase-smadsl
#14
Larissa Romanello, Vitor Hugo Balasco Serrão, Juliana Roberta Torini, Louise E Bird, Joanne E Nettleship, Heather Rada, Yamini Reddivari, Ray J Owens, Ricardo DeMarco, José Brandão-Neto, Humberto D'Muniz Pereira
Schistosoma mansoni is the parasite responsible for schistosomiasis, a disease that affects about 218 million people worldwide. Currently, both direct treatment and disease control initiatives rely on chemotherapy using a single drug, praziquantel. Concerns over the possibility of resistance developing to praziquantel, have stimulated efforts to develop new drugs for the treatment of schistosomiasis. Schistosomes do not have the de novo purine biosynthetic pathway, and instead depend entirely on the purine salvage pathway to supply its need for purines...
March 25, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28322872/plasmodium-hsp40-and-human-hsp70-a-potential-cochaperone-chaperone-complex
#15
Payal Jha, Shyamasree Laskar, Swati Dubey, Mrinal K Bhattacharyya, Sunanda Bhattacharyya
Out of the total forty four members of Plasmodium falciparum Hsp40 protein family, nineteen of them possess a PEXEL motif, and are predicted to be exported into the cytosol of an infected RBC. It is speculated that the human Hsp70 (hHsp70), which resides into the cytosol of the host erythrocyte, along with the exported PfHsp40s assists in the folding of parasitic proteins, thus playing a crucial role in the establishment of virulence. However, till date no experimental evidence supports this hypothesis. Our work establishes that the PEXEL motifs containing Type II PfDNAJ proteins specifically interact with hHsp70 (HSPA1A)...
March 18, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28322871/recurrent-emergence-of-structural-variants-of-ltr-retrotransposon-csrn1-evolving-novel-expression-strategy-and-their-selective-expansion-in-a-carcinogenic-liver-fluke-clonorchis-sinensis
#16
Seon-Hee Kim, Yoon Kong, Young-An Bae
Autonomous retrotransposons, in which replication and transcription are coupled, encode the essential gag and pol genes as a fusion or separate overlapping form(s) that are expressed in single transcripts regulated by a common upstream promoter. The element-specific expression strategies have driven development of relevant translational recoding mechanisms including ribosomal frameshifting to satisfy the protein stoichiometry critical for the assembly of infectious virus-like particles. Retrotransposons with different recoding strategies exhibit a mosaic distribution pattern across the diverse families of reverse transcribing elements, even though their respective distributions are substantially skewed towards certain family groups...
March 18, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28286026/clonorchis-sinensis-excretory-secretory-products-promote-the-migration-and-invasion-of-cholangiocarcinoma-cells-by-activating-the-integrin-%C3%AE-4-fak-src-signaling-pathway
#17
Jhang Ho Pak, Qudsia Bashir, In Ki Kim, Sung-Jong Hong, Sejung Maeng, Young Yil Bahk, Tong-Soo Kim
Cholangiocarcinoma (CCA) is a slow-growing but highly metastatic cancer. Its metastatic potential largely explains its high mortality rate. A recognized risk factor for CCA development is infection with the liver flukes Opisthorchis viverrini and Clonorchis sinensis. We previously reported that the excretory-secretory products (ESPs) of C. sinensis promoted the three-dimensional aggregation and invasion of CCA cells. In the present study, a quantitative real-time PCR array of extracellular matrix (ECM) and adhesion molecules was used to examine the regulatory mechanism of ESP-mediated CCA cell migration and invasion...
March 8, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28274857/bapta-am-decreases-cellular-ph-inhibits-acidocalcisome-acidification-and-autophagy-in-amino-acid-starved-t-brucei
#18
Feng-Jun Li, Kevin S W Tan, Cynthia Y He
To investigate the role of Ca(2+) signaling in starvation-induced autophagy in Trypanosoma brucei, the causative agent of human African trypanosomiasis, we used cell-permeant Ca(2+) chelator BAPTA-AM and cell impermeant chelator EGTA, and examined the potential involvement of several intracellular Ca(2+) signaling pathways in T. brucei autophagy. The results showed an unexpected effect of BAPTA-AM in decreasing cellular pH and inhibiting acidocalcisome acidification in starved cells. The implication of these results in the role of Ca(2+) signaling and cellular/organellar pH in T...
April 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28232060/zinc-finger-nuclease-technology-a-stable-tool-for-high-efficiency-transformation-in-bloodstream-form-t-brucei
#19
Gabriela Schumann, Monica M Kangussu-Marcolino, Nicholas Doiron, Sandro Käser, Gabriela de Assis Burle-Caldas, Wanderson D DaRocha, Santuza M Teixeira, Isabel Roditi
In Trypanosoma brucei, the generation of knockout mutants is relatively easy compared to other organisms as transfection methods are well established. These methods have their limitations, however, when it comes to the generation of genome-wide libraries that require a minimum of several hundred thousand transformants. Double-strand breaks with the meganuclease ISce-I dramatically increase transformation efficiency, but are not widely in use as cell lines need to be generated de novo before each transfection...
April 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28223095/palmitoylation-of-plasmodium-alveolins-promotes-cytoskeletal-function
#20
Annie Z Tremp, Fatimah S Al-Khattaf, Johannes T Dessens
S-palmitoylation is a post-translational lipid modification that is widespread among Plasmodium proteins and essential for parasite development. Little is known about the contribution of palmitoylation to the function of individual parasite molecules and structures. Alveolins are major components of the subpellicular network (SPN), a cortical cytoskeleton primarily involved in providing mechanical strength to the cell. We show here that the alveolin IMC1c is palmitoylated on a conserved cysteine motif, and that non-palmitoylated IMC1c displays normal expression, stability and trafficking...
April 2017: Molecular and Biochemical Parasitology
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