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zenodo44/100

Identification of a lineage-specific protein network at the trypanosome nuclear envelope

<p>The nuclear envelope (NE) separates translation and transcription and is the location of multiple functions, including chromatin organization, nucleocytoplasmic transport, ribosomal maturation and mRNA processing/quality control.&nbsp; The molecular basis for many of these functions have diverged between different eukaryotic lineages.&nbsp; <em>Trypanosoma brucei</em>, a member of the early branching eukaryotic lineage Discoba, highlight many of these, including a distinct lamina and kinetochore composition.&nbsp; Here we describe a cohort of proteins interacting with both the lamina and NPC, which we term lamina-associated proteins (LAPs).&nbsp; LAPs represent a diverse group of proteins, including two candidate NPC-anchoring pore membrane proteins (POMs) with architecture conserved with <em>S. cerevisiae </em>Pom152 and <em>H. sapiens </em>Nup210, and additional peripheral components of the NPC.&nbsp; While many of the LAPs are specific to Trypanosomatids, we also identified broadly conserved proteins, indicating an amalgam of divergence and conservation within the NE proteome of trypanosomes, highlighting the diversity of nuclear biology across the eukaryotes and increasing our understanding of eukaryotic and NPC evolution.</p>

opencc-by-4.0Dec 2023View details →
zenodo40/100

Spatial integration of transcription and splicing in a dedicated compartment sustains monogenic antigen expression in African trypanosomes

<p>This repository contains the data for the manuscript <a href="https://doi.org/10.1038/s41564-020-00833-4">https://doi.org/10.1038/s41564-020-00833-4</a>.</p> <p>The HiC analysis pipeline can be found at&nbsp;<a href="https://github.com/bgbrink/PRJEB35632">https://github.com/bgbrink/PRJEB35632</a>.</p> <p><strong>Abstract</strong></p> <p>Highly selective gene expression is a key requirement for antigenic variation in several pathogens, allowing evasion of host immune responses and maintenance of persistent infections. African trypanosomes &mdash; parasites that cause lethal diseases in humans and livestock &mdash; employ an antigenic variation mechanism that involves monogenic antigen expression from a pool of &gt;2,600 antigen-coding genes. In other eukaryotes, the expression of individual genes can be enhanced by mechanisms involving the juxtaposition of otherwise distal chromosomal loci in the three-dimensional nuclear space. However, trypanosomes lack classical enhancer sequences or regulated transcription initiation. In this context, it has remained unclear how genome architecture contributes to monogenic transcription elongation and transcript processing. Here, we show that the single expressed antigen-coding gene displays a specific inter-chromosomal interaction with a major messenger RNA splicing locus. Chromosome conformation capture (Hi-C) revealed a dynamic reconfiguration of this inter-chromosomal interaction upon activation of another antigen. Super-resolution microscopy showed the interaction to be heritable and splicing dependent. We found a specific association of the two genomic loci with the antigen exclusion complex, whereby VSG exclusion 1 (VEX1) occupied the splicing locus and VEX2 occupied the antigen-coding locus. Following VEX2 depletion, loss of monogenic antigen expres- sion was accompanied by increased interactions between previously silent antigen genes and the splicing locus. Our results reveal a mechanism to ensure monogenic expression, where antigen transcription and messenger RNA splicing occur in a specific nuclear compartment. These findings suggest a new means of post-transcriptional gene regulation.</p>

opencc-by-4.0Jan 2021View details →
zenodo40/100

Fig. 4 in Sexual differences in prevalence of a new species of trypanosome infecting túngara frogs

Fig. 4. Phylogeny of the aquatic clade, and PTP species delimitation results. Best maximum likelihood tree of the18S rRNA gene of member of the aquatic clade and selected outgroups. Numbers on the branches represent support values corresponding to ±70% bootstrap replicates (left) and ±0.9 Bayesian posterior probabilities (right). Subclades are highlighted with colored boxes to indicate host associations. Color of the branches indicate the PTP species delimitation results; monophyletic groups in red indicate members of a single species, blue terminal branches indicate that only one sample is included in such species. Names of the terminals indicate the GenBank accession numbers, scientific name, and sample or isolate code. Star indicates the position of T. tungarae n. sp. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

opencc-by-4.0Apr 2016View details →
zenodo40/100

Fig. 3 in Sexual differences in prevalence of a new species of trypanosome infecting túngara frogs

Fig. 3. Light microscopy of Trypanosoma tungarae n. sp. (Giemasa-staining). (a e) Trypomastigotes stained using Hemacolor ® Giemsa stain kit (Voigt Global Distribution Inc, USA); ‾ (f‾i) Trypomastigotes stained using Giemsa stain following Mohr (1981). Scale bars: 10 µm.

opencc-by-4.0Apr 2016View details →
zenodo40/100

Fig. 1 in Sexual differences in prevalence of a new species of trypanosome infecting túngara frogs

Fig. 1. Photographs of túngara frogs (Engystomops pustulosus) and frog-biting midges (Corethrella spp). (a) Calling male túngara frog preyed upon by frog-biting midges; (b) female (bottom) in amplexus with a male (top) covered with biting midges; (c) female (bottom) with a biting midge on her nostril that was passed from the male during amplexus. Túngara frogs are about 30 mm long while the frog-biting midges are only about 1.5 mm. Photos taken by Alexander Baugh (a) and Ximena E Bernal (b,c).

opencc-by-4.0Apr 2016View details →
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Fig. 2 in Sexual differences in prevalence of a new species of trypanosome infecting túngara frogs

Fig. 2. Map of the Republic of Panaḿa indicating with a star the location of Gamboa, the type locality of Trypanosoma tungarae n. sp. Insert shows the location of Panamáin the New World.

opencc-by-4.0Apr 2016View details →
zenodo40/100

Fig. 2 in Phlebotomine Sandflies - Potential Vectors of Avian Trypanosomes

Fig. 2. Light microscopy of live T. avium SIM3 trypanosomes in L. longipalpis (A) and P. arabicus (B) gut; Giemsa stained metacyclic trypomastigotes of T. avium (s. s.) strain SIM3 from L. longipalpis hindgut (C). Scanning electron microscopy of T. avium (s. s.) SIM3 in the gut of sandfly P. arabicus (D, E) and L. longipalpis (F). A detailed view of trypomastigotes at the epithelium (D); note the massive infection of trypomastigots covering the gut (E); rounded objects are developmental stages of gregarines Psychodiella chagasi (F).

opencc-by-4.0Dec 2018View details →
zenodo40/100

Fig 1 in Phlebotomine Sandflies - Potential Vectors of Avian Trypanosomes

Fig 1. Infection rates and intensities in sandflies L. longipalpis and P. arabicus membrane-fed on Trypanosoma avium strains BUT15 and SIM3, and on canaries. Infection intensities: low – 1–100 parasites; medium – 100–1000 parasites; heavy –&gt; 1000 parasites per gut. Numbers of dissected females shown above the columns. Data from repeated experimental feeding of L. longipalpis on SIM3 are pooled. Sandflies fed on membrane were dissected after 8 days, L. longipalpis fed on canaries were dissected 7–10 days after feeding.

opencc-by-4.0Dec 2018View details →
zenodo40/100

Fig. 11 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 11. Transmission electron micrograph of an epimastigote and an amastigote of G2 (Clade A). (A) Epimastigote in culture; Ax: Axoneme showing nine doublets of microtubules surrounding a central pair; Ac: Acidocalcisomes; Arrow: Subpellicular microtubules. (B) Amastigote inside a VERO cell. Scale bars = 0.5 µm (A), 1 µm (B).

opencc-by-4.0Dec 2013View details →
zenodo40/100

Fig. 9 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 9. Infection of Vero (A) and L6 cells (B) with G2 (Clade A) and T. cruzi as a positive control of infection (Diff-Quick stained). (A) Intracellular amastigotes of G2. (B) Intracellular amastigotes of T. cruzi. Scale bars = 10 µm.

opencc-by-4.0Dec 2013View details →
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Fig. 8 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 8. Epimastigotes of G1 and G2 (Clade A) arranged in rosettes in culture. (A) Diff-Quick stained rosettes. (B) Rosettes in fresh wet preparations showing numerous intracellular acidocalcisomes. Scale bars = 10 µm.

opencc-by-4.0Dec 2013View details →
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Fig. 7 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 7. Light microscopy of Diff-Quick stained blood and culture forms of G1 and G2 (Clade A) (A) Trypomastigote in blood of a woylie naturally infected; (B) slender epimastigote in culture; (C and D) shaped epimastigote in culture; (E) spheromastigote in culture; (F) spheromastigotes dividing in culture. Scale bars = 10 µm.

opencc-by-4.0Dec 2013View details →
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Fig. 6 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 6. Structures suggestive of amastigotes (arrows) of G2 (Clade A) in heart tissue positive by PCR (H&amp;E stained). Scale bars = (A) 20 µm, (B) 10 µm.

opencc-by-4.0Dec 2013View details →
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Fig. 5 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 5. Histopathology of two woylies naturally infected with G2 (Clade A) (H&amp;E stained). (A) Multifocal, moderate to severe, chronic, pyogranulomatous myocarditis and (B) endocarditis. (C) Mineralisation of heart tissue. (D) Tongue showing multifocal, moderate, chronic, pyogranulomatous glossitis. (E) Skeletal muscle degeneration. (F) Inflammatory cells around a blood vessel. Scale bars = 20 µm.

opencc-by-4.0Dec 2013View details →
zenodo40/100

Fig. 3 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 3. Phylogenetic relationships of the new trypanosome isolates from Western Australian marsupials based on gGAPDH sequences (~810 bp) using Mr Bayes. The tree was rooted with five sequences as outgroups. Bayesian posterior probabilities are shown at nodes. Bar, 0.07 substitutions per site.

opencc-by-4.0Dec 2013View details →
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Fig. 10 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 10. Scanning electron micrograph of G2 (Clade A) grown in culture with Vero cells. (A) Trypomastigote invading a cell, with the flagella still external to the cell. (B) Dead cell(s) surrounded by amastigotes and trypomastigotes. Scale bars = 2 µm (A), 4 µm (B).

opencc-by-4.0Dec 2013View details →
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Fig. 4 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 4. Prevalence of infection with trypanosomes within the different clades in woylies from the stable and declining populations. 95% confidence intervals (95% CI).

opencc-by-4.0Dec 2013View details →
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Fig. 2 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 2. Phylogenetic analysis of the relationships between Australian trypanosomes based on 18S rDNA sequences. Phylogenetic trees were constructed by the Bayesian method sequences (~1410 bp). (B) Phylogenetic position of shorter 18S rDNA sequences (786 bp) CHA1, TRY1, TRY2, WYA1, WYA2, BDA1, Q3, Q10, GP63 and GP94. Threes were rooted with from Bayesian posterior probabilities are shown at nodes. In red: trypanosome genotypes found in this study. Bar, 0.2 substitutions per site.

opencc-by-4.0Dec 2013View details →
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Fig. 2. A in Trypanosomes of Australian mammals: A review

Fig. 2. A graphical representation of the phylogenetic relationship shared by some Australian trypanosome isolates based on gGAPDH sequences (=810 bp) (reproduced with permission from Botero et al. (2013), with modifications highlighted in grey).

opencc-by-4.0Aug 2014View details →
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Fig. 1 in Trypanosomes of Australian mammals: A review

Fig. 1. (a) General trypanosome shape (trypomastigote form from the blood of a woylie (Bettongia penicillata)) K = kinetoplast, N = nucleus and FF = free flagellum and RBC = red blood cells, (b) host: woylie (Bettongia penicillata).

opencc-by-4.0Aug 2014View details →

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