Trans-splicing of leader sequences onto the 5'ends mRNAs is a widespread phenomenon in protozoa, nematodes and some chordates. Using parallel sequencing we have developed method to simultaneously map 5'splice sites analyze corresponding gene expression profile, that term spliced trapping (SLT). The can be applied any organism with sequenced genome trans-splicing conserved sequence. We analyzed profiles splicing patterns bloodstream insect forms parasite Trypanosoma brucei. detected 5' splice...

Abstract Background Trypanosoma brucei is the causative agent of human African sleeping sickness and Nagana in cattle. In addition to being an important pathogen T . has developed into a model system cell biology. Results Using Stable Isotope Labelling Amino acids Cell culture (SILAC) combination with mass spectrometry we determined abundance >1600 proteins long slender (LS), short stumpy (SS) mammalian bloodstream form stages relative procyclic (PC) insect-form stage. total identified...

Abstract The intracellular ATP-ribosyltransferases PARP1 and PARP2, contribute to DNA base excision repair (BER) demethylation have been implicated in epigenetic programming early mammalian development. Recently, proteomic analyses identified BER proteins be covalently poly-ADP-ribosylated by PARPs. role of this posttranslational modification the process is unknown. Here, we show that senses AP-sites SSBs generated during TET-TDG mediated active attaches PAR each protein engaged. Covalent...

Theileria annulata is a tick-transmitted apicomplexan parasite that gained the unique ability among parasitic eukaryotes to transform its host cell, inducing fatal cancer-like disease in cattle. Understanding mechanistic interplay between cell and malignant species drives this transformation requires identification of responsible effector proteins. In study, we used TurboID-based proximity labeling, which unbiasedly identified secreted proteins within compartments. By fusing TurboID nuclear...

The protozoan pathogen Trypanosoma brucei is transmitted between mammals by tsetse flies. first compartment colonised trypanosomes after a blood meal the fly midgut lumen. Trypanosomes present in lumen—designated as early procyclic forms—express stage-specific surface glycoproteins EP and GPEET procyclin. When establish mature infection colonise ectoperitrophic space, down-regulated, becomes major protein of late forms. A few years ago, it was discovered that form exhibit social motility...

It has long been known that trypanosomes regulate mitochondrial biogenesis during the life cycle of parasite; however, protein inventory (MitoCarta) and its regulation remain unknown. We present a novel computational method for genome-wide prediction proteins using support vector machine-based classifier with ∼90% accuracy. Using this method, we predicted localization 468 high confidence have experimentally verified subset these proteins. then applied recently developed parallel sequencing...

AbstractUnlike other eukaryotes, the protein-coding genes of Trypanosoma cruzi are arranged in large polycistronic gene clusters transcribed by polymerase II (Pol II). Thus, it is thought that trypanosomes rely solely on posttranscriptional processes to regulate expression. Here, we show glucosylated thymine DNA base (β-d-glucosyl-hydroxymethyluracil or J) present within sequences flanking units (PTUs) T. cruzi. The loss J at sites transcription initiation, via deletion two enzymes synthesis...

TbRRM1 of Trypanosoma brucei is a nucleoprotein that was previously identified in search for splicing factors T. brucei. We show associates with mRNAs and the auxiliary factor polypyrimidine tract-binding protein 2, but not components core spliceosome. also interacts several retrotransposon hot spot (RHS) proteins histones. RNA immunoprecipitation tagged form from procyclic (insect) trypanosomes ca. 1,500 transcripts were enriched 3,000 underrepresented compared to cellular mRNA. Enriched...

Abstract Mice deficient for ADP-ribosyltransferase diphteria toxin–like 1 (ARTD1) are protected against microbially induced inflammation. To address the contribution of ARTD1 to inflammation specifically in myeloid cells, we generated an Artd1ΔMyel mouse strain with conditional deficiency lineages and examined three disease models. We found that ARTD1, but not its enzymatic activity, enhanced transcriptional activation distinct LPS-induced genes included IL-12, TNF-α, IL-6 primary bone...

ABSTRACT Induction of nuclear factor activated T cell cytoplasmic 1 (NFATc1) by macrophage colony-stimulating (M-CSF) and receptor activator NF-κB ligand (RANKL) is essential for differentiation into osteoclasts (OCs), but the underlying mechanisms remain unclear. The ability poly(ADP-ribose) polymerase (PARP1) to poly-ADP-ribosylate NFATc1 in cells prompted us investigate PARP1 interaction during osteoclastogenesis. However, extensive studies failed directly link NFATc1. A combination...

The spike protein VP4 participates in diverse steps of the rotavirus (RV) life cycle, including virus-cell attachment, internalization, modulation endocytosis, virion morphogenesis, and virus egress. Using reverse genetics, we constructed for first time a recombinant RV, rRV/VP4-BAP, harboring heterologous peptide lectin domain (loop K145-G150) VP4.

During degeneration of the intervertebral disc ingrowth blood vessels and nerves into are associated with back pain. Vascular endothelial growth factors promote vasculogenesis by binding to membrane vascular factor receptor 1, while shorter soluble forms this can inhibit vascularization. We hypothesized that might change between stages degeneration. Expression 1 in human degenerated discs healthy bovine caudal was assessed qRT-PCR immunoblot. Comparative microarray meta-analysis across...

Sterols are an essential class of lipids in eukaryotes, where they serve as structural components membranes and play important roles signaling molecules. also high pharmacological significance: cholesterol-lowering drugs blockbusters human health, inhibitors ergosterol biosynthesis widely used antifungals. Inhibitors synthesis being developed for Chagas's disease, caused by Trypanosoma cruzi. Here we develop silico pipeline to globally evaluate sterol metabolism perform comparative genomics....

While protein ADP-ribosylation was reported to regulate differentiation and dedifferentiation, it has so far not been studied during transdifferentiation. Here, we found that MyoD-induced transdifferentiation of fibroblasts myoblasts promotes the expression ADP-ribosyltransferase ARTD1. Comprehensive analysis genome architecture by Hi-C RNA-seq indicated ARTD1 locally contributed A/B compartmentalization coregulated a subset MyoD target genes were however sufficient alter Surprisingly,...

Abstract The formation of viroplasms is a well-conserved step in the rotavirus (RV) life cycle. In these structures, both virus genome replication and progeny assembly take place. A stabilized microtubule cytoskeleton lipid droplets are required for viroplasm formation, which involves several proteins. viral spike protein VP4 has not previously been shown to have direct role formation. However, it involved with virus-cell attachment, endocytic internalization, virion morphogenesis. Moreover,...

Abstract Background/Aims Giant-cell arteritis (GCA) is a large vessel vasculitis predominantly affecting those above 50 years of age. Creutzfeldt-Jacob disease (CJD) not well-recognised differential diagnosis GCA presentation. Prion diseases are progressive, currently untreatable and ultimately fatal. Methods We report the case 76-year-old female who had an eventful hospital stay 37 days. She has history asthma, hypertension, ulcerative colitis right-sided 5 back (normal inflammatory markers...

ABSTRACT Theileria annulata is a tick-transmitted apicomplexan parasite that gained the unique ability among parasitic eukaryotes to transform its host cell, inducing fatal cancer-like disease in cattle. Understanding mechanistic interplay driving this transformation between cell and malignant species requires identification of responsible effector proteins. In study, we used TurboID-based proximity labelling, which unbiasedly identified secreted proteins within compartments. By fusing...