A5080164627- Trypanosoma species research and implications
- CRISPR and Genetic Engineering
- Mosquito-borne diseases and control
- RNA regulation and disease
- RNA and protein synthesis mechanisms
- Malaria Research and Control
- Viral Infections and Immunology Research
- Invertebrate Immune Response Mechanisms
- RNA Research and Splicing
- Biochemical and Molecular Research
- Insect symbiosis and bacterial influences
- RNA modifications and cancer
- Research on Leishmaniasis Studies
- Genomics and Chromatin Dynamics
- Viral Infections and Vectors
- Peptidase Inhibition and Analysis
- Silk-based biomaterials and applications
- Developmental Biology and Gene Regulation
- interferon and immune responses
- Superconductivity in MgB2 and Alloys
- Neurobiology and Insect Physiology Research
- Epigenetics and DNA Methylation
- Muscle Physiology and Disorders
- Chromatin Remodeling and Cancer
- Genetics and Neurodevelopmental Disorders
Infectious Disease Research Institute
2015-2024
University of Washington
2022-2024
Seattle Children's Hospital
2022
Center for Infectious Disease Research
2012-2019
Seattle University
2015
University of Oxford
2012-2014
University of Edinburgh
2005
Wellcome Centre for Cell Biology
2005
Two key biological features distinguish Trypanosoma evansi from the T. brucei group: independence tsetse fly as obligatory vector, and need for functional mitochondrial DNA (kinetoplast or kDNA). In an effort to better understand molecular causes consequences of these differences, we sequenced genome akinetoplastic strain China compared it b. reference strain. The annotated shows extensive similarity reference, with 94.9% predicted coding sequences (CDS) having ortholog in evansi, 94.6%...
Summary In the Drosophila oocyte, mRNA transport and localised translation play a fundamental role in axis determination germline formation of future embryo. gurken encodes secreted TGF-α signal that specifies dorsal structures, is to dorso-anterior corner oocyte via cis-acting 64 nucleotide localisation signal. Using GRNA chromatography, we characterised biochemical composition ribonucleoprotein complexes form around oocyte. We identified number factors already known be involved...
Localized mRNA translation is thought to play a key role in synaptic plasticity, but the identity of transcripts and molecular mechanism underlying their function are still poorly understood. Here, we show that Syncrip, regulator localized Drosophila oocyte component mammalian neuronal granules, also expressed larval neuromuscular junction, where it regulates growth. We use RNA-immunoprecipitation followed by high-throughput sequencing qRT-PCR Syncrip associates with number mRNAs encoding...
U-insertion/deletion (U-indel) RNA editing in trypanosome mitochondria is directed by guide RNAs (gRNAs). This may developmentally control respiration bloodstream forms (BSF) and insect procyclic (PCF). Holo-editosomes include the accessory Editing Substrate Binding Complex (RESC) Helicase 2 (REH2C), but specific proteins controlling differential remain unknown. Also, appears highly error prone because most U-indels do not match canonical pattern. However, despite extensive non-canonical of...
RNA editing is an essential post-transcriptional process that creates functional mitochondrial mRNAs in Kinetoplastids. Multiprotein editosomes catalyze pre-mRNA cleavage, uridine (U) insertion or deletion, and ligation as specified by guide RNAs. Three functionally compositionally distinct differ the mutually exclusive presence of KREN1, KREN2 KREN3 endonuclease their associated partner proteins. Because cleavage a likely point regulation for editing, we elucidated specificity vivo. We used...
Significance Trypanosoma brucei is a deadly kinetoplastid parasite that causes the human and veterinarian diseases African sleeping sickness nagana. We combine chemical cross-linking mass spectrometry, structural modeling, genetics, biochemistry to define global architecture of RNA editing “editosome” complexes in these parasites. Editosomes are unique kinetoplastids, which also include cruzi Leishmania parasites cause Chagas disease Leishmaniases, respectively. essential for viability...
The transcriptome of kinetoplastid mitochondria undergoes extensive RNA editing that inserts and deletes uridine residues (U's) to produce mature mRNAs. editosome is a multiprotein complex provides endonuclease, TUTase, exonuclease, ligase activities required for editing. editosome's KREPB4 KREPB5 proteins are essential integrity parasite viability contain semi-conserved motifs corresponding zinc finger, RNase III, PUF domains, but date no functional analysis these domains has been reported....
The invasion of a suitable host hepatocyte by Plasmodium sporozoites is an essential step in malaria infection. We demonstrate that infected hepatocytes, lysosomes are redistributed away from the nucleus, and surface exposure lysosome-associated membrane protein 1 (LAMP1) increased. Lysosome exocytosis cells occurs independently sporozoite traversal. Instead, sporozoite-secreted factor sufficient for process. Knockdown SNARE proteins involved lysosome-plasma fusion reduces lysosome In...
Understanding immune mechanisms that mediate malaria protection is critical for improving vaccine development. Vaccination with radiation-attenuated Plasmodium falciparum sporozoites (PfRAS) induces high level of sterilizing immunity against and serves as a valuable tool the study protective mechanisms. To identify vaccine-induced protection-associated responses during malarial infection, we performed transcriptome profiling whole blood in-depth cellular PBMCs from volunteers who received...
Immunization with radiation-attenuated sporozoites (RAS) can confer sterilizing protection against malaria, although the mechanisms behind this are incompletely understood. We performed a systems biology analysis of samples from by Mosquito Radiation Attenuated Sporozoites (IMRAS) trial, which comprised P. falciparum RAS-immunized (PfRAS), malaria-naive participants whose malaria infection was subsequently assessed controlled human (CHMI). Blood collected after initial PfRAS immunization...
KREPB5 is an essential component of ϳ20S editosomes in Trypanosoma brucei which contains a degenerate, noncatalytic RNase III domain.To explore the function this protein, we used novel approach to make and screen numerous conditional null T. bloodstream form cell lines that express randomly mutagenized alleles.We identified nine single amino acid substitutions could not complement loss wild-type KREPB5.Seven these were within domain, two C-terminal region has no homology known...
Uridine insertion and deletion RNA editing generates functional mitochondrial mRNAs in Trypanosoma brucei, several transcripts are differentially edited bloodstream (BF) procyclic form (PF) cells correlating with changes function. Editing is catalyzed by three ∼20S editosomes that have a common set of 12 proteins, but typified mutually exclusive RNase III KREN1, N2, N3 endonucleases distinct cleavage specificities. KREPB4 editosome protein has degenerate domain lacking conserved catalytic...
In the Drosophila oocyte, gurken (grk) mRNA encodes a secreted TGF-α signal that specifies future embryonic dorso-ventral axes by altering fate of surrounding epithelial follicle cells. We previously identified number RNA binding proteins associate specifically with 64 nucleotide grk localization signal, including orthologue polypyrimidine tract-binding protein (PTB), Hephaestus (Heph). To test whether Heph is required for correct or function, we used immunoprecipitation to validate...
Multiprotein editosomes catalyze gRNA-specified insertion and deletion of uridines to create functional mitochondrial mRNAs in Trypanosoma brucei . Three functionally distinct are distinguished by their single KREN1, KREN2, or KREN3 RNase III endonuclease and, respectively, KREPB8, KREPB7, KREPB6 partner proteins. These endonucleases perform the first catalytic step editing, cleaving mRNA diverse mRNA/gRNA heteroduplex substrates. We identified divergent likely noncatalytic domains KREPB4,...
Kinetoplastid pathogens including Trypanosoma brucei , T. cruzi and Leishmania species, are early diverged, eukaryotic, unicellular parasites. Functional understanding of many proteins from these has been hampered by limited sequence homology to other model organisms. Here we describe the development a high-throughput deep mutational scanning approach in that facilitates rapid unbiased assessment impacts possible amino acid substitutions within protein on cell fitness, as measured relative...
Abstract Unknown factors regulate mitochondrial U-insertion/deletion (U-indel) RNA editing in procyclic-form (PCF) and bloodstream-form (BSF) T. brucei. This editing, directed by anti-sense gRNAs, creates canonical protein-encoding mRNAs may developmentally control respiration. Canonical gRNAs that specify mRNA sequences occurs amid massive non-canonical of unclear sources biological significance. We found PCF-specific repression at a major early checkpoint ND7, involving helicase...
Identifying immune processes required for liver-stage sterilizing immunity to malaria remains an open problem. The IMRAS trial comprised 5x immunizations with radiation-attenuated sporozoites resulting in 55% protection from subsequent challenge. To identify correlates of vaccination and protection, we performed detailed systems immunology longitudinal profiling the entire time course including whole blood transcriptomics, PBMC cell phenotyping serum antigen array 11 sporozoite (RAS)...
Trypanosoma and Leishmania parasites cause devastating tropical diseases resulting in serious global health consequences. These organisms have complex life cycles with mammalian hosts insect vectors. The must, therefore, survive different environments, demanding rapid physiological metabolic changes. responses depend upon regulation of gene expression, which primarily occurs posttranscriptionally. Altering the composition or conformation RNA through nucleotide modifications is one...
Mitochondrial gene expression in trypanosomes requires numerous multiprotein complexes that are unique to kinetoplastids. Among these, the most well characterized RNA editing catalytic (RECCs) catalyze guide (gRNA)-specified insertion and deletion of uridines during mitochondrial mRNA maturation. This post-transcriptional resequencing mRNAs can be extensive, involving dozens different gRNAs hundreds sites with mature sequences resulting from process. Proper coordination cognate is attributed...
Editosomes are the multiprotein complexes that catalyze insertion and deletion of uridines to create translatable mRNAs in mitochondria kinetoplastids. Recognition cleavage a broad diversity RNA substrates vivo require three functionally distinct RNase III-type endonucleases, as well five additional editosome proteins contain noncatalytic III domains. domains have recently been identified accessory KREPB9 KREPB10, suggesting role related editing endonuclease function. In this report, we...
Trypanosoma brucei and related kinetoplastid parasites possess unique RNA processing pathways, including in their mitochondria, that regulate metabolism development. Altering composition or conformation through nucleotide modifications is one such pathway, pseudouridine fate function many organisms. We surveyed synthase (PUS) orthologs trypanosomatids, with a particular interest mitochondrial enzymes due to potential importance for metabolism. (mt)-LAF3 an ortholog of human yeast PUS...
The gRNA directed U-insertion and deletion editing of mitochondrial mRNAs that is essential in different life-cycle stages for the protozoan parasite Trypanosoma brucei performed by three similar multiprotein catalytic complexes (CCs) contain requisite enzymes. These CCs also a common set eight proteins have no apparent direct function, including six an OB-fold domain. We show here one these proteins, KREPA3 (A3), has structural homology to other editing, multifunctional. investigated A3...