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Molecular and Biochemical Parasitology

Mridula Agrahari, Amit Kumar Gaurav, Alok Bhattacharya, Sudha Bhattacharya
Non LTR retrotransposons (EhLINEs and EhSINEs) occupy 11% of the Entamoeba histolytica genome. Since promoter DNA methylation at cytosines has been correlated with transcriptional silencing of transposable elements in model organisms we checked whether this was the case in EhLINE1. We located promoter activity in a 841bp fragment at 5'-end of this element by luciferase reporter assay. From RNAseq and RT-PCR analyses we selected a transcriptionally active and silent copy to study cytosine DNA methylation of the promoter region by bisulfite sequencing...
January 9, 2017: Molecular and Biochemical Parasitology
Olivera Cirovic, Roman Trikin, Anneliese Hoffmann, Nicholas Doiron, Martin Jakob, Torsten Ochsenreiter
RNA recognition motif (RRM) containing proteins are important regulators of gene expression in trypanosomes. Here we expand our current knowledge on the exclusively nuclear localized RRM domain containing protein RBP33 of Trypanosoma brucei. Overexpression of RBP33 leads to a quick growth arrest in G2/M in bloodstream form cells likely due to an overall mRNA- and spliced leader abundance decrease while the ribosomal RNAs remain unaffected. The recombinant RBP33 binds to poly(A) and random sequence RNA in vitro confirming its role as a RNA binding protein...
January 6, 2017: Molecular and Biochemical Parasitology
Jose M Requena, Alberto Rastrojo, Esther Garde, Manuel C López, M Carmen Thomas, Begoña Aguado
The genomes of most eukaryotic organisms contain a large number of transposable elements that are able to move from one genomic site to another either by transferring of DNA mobile elements (transposons) or transpose via reverse transcription of an RNA intermediate (retroposons). An exception to this rule is found in protists of the subgenus Leishmania, in which active retroposons degenerated after a flourishing era, leaving only retroposon remains; these have been classified into two families: SIDER1 and SIDER2...
December 27, 2016: Molecular and Biochemical Parasitology
Cui Zhang, Han Gao, Zhenke Yang, Yuanyuan Jiang, Zhenkui Li, Xu Wang, Bo Xiao, Xin-Zhuan Su, Huiting Cui, Jing Yuan
CRISPR/Cas9 has been successfully adapted for gene editing in malaria parasites including Plasmodium falciparum and Plasmodium yoelii. However, the reported methods were limited to editing one gene at a time. In practice, it is often desired to modify multiple genetic loci in a parasite genome. Here we describe a CRISPR/Cas9 mediated genome editing method that allows successive modification of more than one gene in the genome of P. yoelii using an improved single-vector system (pYCm) we developed previously...
December 26, 2016: Molecular and Biochemical Parasitology
Akram A Da'dara, Giles Siddons, Melissa Icaza, Qiang Wang, Patrick J Skelly
Schistosomes are intravascular parasitic worms that cause the debilitating disease schistosomiasis. To better understand how these long-lived parasites may subvert host immune and hemostatic capabilities, we examine here the impact of adult Schistosoma mansoni worms on the human serum proteome. Normal human serum (150μl) was incubated at 37°C for one hour either in the presence or absence of adult worms (∼50 pairs). Thereafter parasites were removed, serum samples were labeled and their proteins resolved for comparative analysis by 2D-Differential in-Gel Electrophoresis (2D-DIGE)...
December 20, 2016: Molecular and Biochemical Parasitology
Cornelis H Hokke, Angela van Diepen
Glycoproteins and glycolipids of parasitic helminths play important roles in biology and host-parasite interaction. This review discusses recent helminth glycomics studies that have been expanding our insights into the glycan repertoire of helminths. Structural data are integrated with biological and immunological observations to highlight how glycomics advances our understanding of the critical roles that glycans and glycan motifs play in helminth infection biology. Prospects and challenges in helminth glycomics and glycobiology are discussed...
December 6, 2016: Molecular and Biochemical Parasitology
Kevin L Howe, Bruce J Bolt, Myriam Shafie, Paul Kersey, Matthew Berriman
The number of publicly available parasitic worm genome sequences has increased dramatically in the past three years, and research interest in helminth functional genomics is now quickly gathering pace in response to the foundation that has been laid by these collective efforts. A systematic approach to the organisation, curation, analysis and presentation of these data is clearly vital for maximising the utility of these data to researchers. We have developed a portal called WormBase ParaSite (http://parasite...
November 27, 2016: Molecular and Biochemical Parasitology
Jonathan D Stoltzfus, Adeiye A Pilgrim, De'Broski R Herbert
The advent of high-throughput, next-generation sequencing methods combined with advances in computational biology and bioinformatics have greatly accelerated discovery within biomedical research. This "post-genomics" era has ushered in powerful approaches allowing one to quantify RNA transcript and protein abundance for every gene in the genome - often for multiple conditions. Herein, we chronicle how the post-genomics era has advanced our overall understanding of parasitic nematodes through transcriptomics and proteomics and highlight some of the important advances made in each major nematode clade...
November 22, 2016: Molecular and Biochemical Parasitology
Paula Arbildi, Silvana La-Rocca, Veronica Lopez, Natalia Da-Costa, Veronica Fernandez
In the cestode parasite Echinococcus granulosus, three phylogenetically distant cytosolic glutathione transferases (GSTs) (EgGST1, 2 and 3) were identified. Interestingly, the C-terminal domains of EgGST3 and EgGST2 but not EgGST1, exhibit all amino acids involved in Sigma-class GST dimerization. Here, we provide evidence indicating that EgGST2 and EgGST3 naturally form a heterodimeric structure (EgGST2-3), and also we report the enzymatic activity of the recombinant heterodimer. EgGST2-3 might display novel properties able to influence the infection establishment...
January 2017: Molecular and Biochemical Parasitology
Amir Zaidi, Krishn Pratap Singh, Vahab Ali
Parasites of genus Leishmania are the causative agents of complex neglected diseases called leishmaniasis and continue to be a significant health concern globally. Iron is a vital nutritional requirement for virtually all organisms, including pathogenic trypanosomatid parasites, and plays a crucial role in many facets of cellular metabolism as a cofactor of several enzymes. Iron acquisition is essential for the survival of parasites. Yet parasites are also vulnerable to the toxicity of iron and reactive oxygen species...
January 2017: Molecular and Biochemical Parasitology
Rémi Longuespée, Rita Casadonte, Mark Kriegsmann, Petra Wandernoth, Katharina Lisenko, Gabriel Mazzucchelli, Michael Becker, Jörg Kriegsmann
Cystic echinococcosis (CE) is a pandemic infectious disease caused by the tapeworm Echinococcus granulosus that forms cysts in different organs such as lungs and liver. Imaging examination and serological tests have some drawbacks such as low sensitivity. In this study, we used an up-to-date workflow of laser microdissection-based microproteomics and matrix-assisted laser desorption/ionization time-of-flight imaging mass spectrometry in order to depict the proteomic pattern of CE in the liver. This investigation revealed specific markers of a parasitic cyst in liver...
January 2017: Molecular and Biochemical Parasitology
Leticia Pérez-Díaz, Tais Caroline Silva, Santuza M R Teixeira
Amastins are surface glycoproteins, first identified in amastigotes of T. cruzi but later found to be expressed in several Leishmania species, as well as in T. cruzi epimastigotes. Amastins are encoded by a diverse gene family that can be grouped into four subfamilies named α, β, γ, and δ amastins. Differential expression of amastin genes results from regulatory mechanisms involving changes in mRNA stability and/or translational control. Although distinct regulatory elements were identified in the 3' UTR of T...
January 2017: Molecular and Biochemical Parasitology
Daniel Musikant, Gabriel Ferri, Ignacio M Durante, Carlos A Buscaglia, Daniel L Altschuler, Martin M Edreira
Mechanistic details of the modulation by cAMP of Trypanosoma cruzi host cell invasion remain ill-defined. Here we report that activation of host's Epac1 stimulated invasion, whereas specific pharmacological inhibition or maneuvers that alter Epac1 subcellular localization significantly reduced invasion. Furthermore, while specific activation of host cell PKA showed no effect, its inhibition resulted in an increased invasion, revealing a crosstalk between the PKA and Epac signaling pathways during the process of invasion...
January 2017: Molecular and Biochemical Parasitology
Amilcar Arenal, Arnielis Díaz, Lew Kan Sprenger, Andreia Buzatti, Maria Angela Machado Fernandes, Jeannie Nascimento Dos Santos, Marcelo Molento
RNA interference has been widely used in parasites. In Haemonchus contortus, reproducible silencing has been reported; however, in this species an altered phenotype has been observed for only a few genes silenced by RNA interference. The aim of this work was to evaluate the effect of silencing Dim-1, a member of the disorganized muscle family on third stage larvae (L3) of H. contortus. Effective silencing of Dim-1 in L3 led to reduced L3 migration and slowed larval development from L3 to early L4. To our knowledge, this is the first report to demonstrate Dim-1 silencing affecting phenotypic characteristics of this parasite...
January 2017: Molecular and Biochemical Parasitology
Smriti Kala, Vaibhav Mehta, Chun Wai Yip, Houtan Moshiri, Hamed Shateri Najafabadi, Ruoyu Ma, Najmeh Nikpour, Sara L Zimmer, Reza Salavati
Ribosomal RNA maturation is best understood in yeast. While substantial efforts have been made to explore parts of these essential pathways in animals, the similarities and uniquenesses of rRNA maturation factors in non-Opisthokonts remain largely unexplored. Eukaryotic ribosome synthesis requires the coordinated activities of hundreds of Assembly Factors (AFs) that transiently associate with pre-ribosomes, many of which are essential. Pno1 and Nob1 are two of six AFs that are required for the cytoplasmic maturation of the 20S pre-rRNA to 18S rRNA in yeast where it has been almost exclusively analyzed...
January 2017: Molecular and Biochemical Parasitology
Amit Kumar Gaurav, Jitender Kumar, Mridula Agrahari, Alok Bhattacharya, Vijay Pal Yadav, Sudha Bhattacharya
Retrotransposons are mobile genetic elements found in most organisms. Their origin and evolution is not very well understood. Retrotransposons that lack long terminal repeats (non-LTR) have been classified based on their reverse transcriptase (RT) and endonuclease sequences into groups, of which R2 is the most ancient. Its members contain a single open reading frame (ORF) while there are two ORFs in the other groups, of which ORF2 contains the RT and endonuclease sequences. It is thought that ORF1 was added later to the single-ORF-containing elements, and codes for a protein with nucleic acid binding activity...
January 2017: Molecular and Biochemical Parasitology
Leobarda Robles-Martinez, Tavis L Mendez, Jennifer Apodaca, Siddhartha Das
The stage differentiation from trophozoite to cyst (i.e., encystation) is an essential step for Giardia to survive outside its human host and spread the infection via the fecal-oral route. We have previously shown that Giardia expresses glucosylceramide transferase 1 (GlcT1) enzyme, the activity of which is elevated during encystation. We have also reported that blocking the activity of gGlcT1 interferes with the biogenesis of encystation-specific vesicles (ESVs) and cyst viability in Giardia. To further understand the role of this enzyme and how it regulates encystation, we overexpressed, knocked down, and rescued the giardial GlcT1 (gGlcT1) gene and measured its enzymatic activity in live parasites as well as in isolated membrane fractions using NBD-ceramide and UDP-glucose or UDP-galactose...
January 2017: Molecular and Biochemical Parasitology
Karin Blomqvist, Christen DiPetrillo, Vincent A Streva, Stewart Pine, Jeffrey D Dvorin
Emerging resistance to current anti-malarials necessitates a more detailed understanding of the biological processes of Plasmodium falciparum proliferation, thus allowing identification of new drug targets. The well-conserved protein Receptor for Activated C-Kinase 1 (RACK1) was originally identified in mammalian cells as an anchoring protein for protein kinase C (PKC) and has since been shown to be important for cell migration, cytokinesis, transcription, epigenetics, and protein translation. The P. falciparum ortholog, PfRACK1, is expressed in blood stages of the parasite and is diffusely localized in the parasite cytoplasm...
January 2017: Molecular and Biochemical Parasitology
Sachin V Surve, Bryan C Jensen, Meredith Heestand, Muriel Mazet, Terry K Smith, Frédéric Bringaud, Marilyn Parsons, Achim Schnaufer
In the slender bloodstream form, Trypanosoma brucei mitochondria are repressed for many functions. Multiple components of mitochondrial complex I, NADH:ubiquinone oxidoreductase, are expressed in this stage, but electron transfer through complex I is not essential. Here we investigate the role of the parasite's second NADH:ubiquinone oxidoreductase, NDH2, which is composed of a single subunit that also localizes to the mitochondrion. While inducible knockdown of NDH2 had a modest growth effect in bloodstream forms, NDH2 null mutants, as well as inducible knockdowns in a complex I deficient background, showed a greater reduction in growth...
January 2017: Molecular and Biochemical Parasitology
Fatimah S Al-Khattaf, Annie Z Tremp, Amira El-Houderi, Johannes T Dessens
Apicomplexan parasites possess a unique cortical cytoskeleton structure composed of intermediate filaments. Its building blocks are provided by a conserved family of proteins named alveolins. The core alveolin structure is made up of tandem repeat sequences, thought to be responsible for the filamentous properties of these proteins. A subset of alveolins also possess conserved motifs composed of three closely spaced cysteine residues situated near the ends of the polypeptides. The roles of these cysteine motifs and their contribution to alveolin function remains poorly understood...
January 2017: Molecular and Biochemical Parasitology
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