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Proteomic and transcriptomic analyses of early and late-chronic Toxoplasma gondii infection shows novel and stage specific transcripts
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Proteomic and transcriptomic analyses of early and late-chronic Toxoplasma gondii infection shows novel and stage specific transcripts
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Journal Article
Proteomic and transcriptomic analyses of early and late-chronic Toxoplasma gondii infection shows novel and stage specific transcripts
2019
Overview
Background
The protozoan pathogen
Toxoplasma gondii
has the unique ability to develop a chronic infection in the brain of its host by transitioning from the fast growing tachyzoite morphology to latent bradyzoite morphology. A hallmark of the bradyzoite is the development of neuronal cysts that are resilient against host immune response and current therapeutics. The bradyzoite parasites within the cyst have a carbohydrate and protein-rich wall and a slow-replication cycle, allowing them to remain hidden from the host. The intracellular, encysted lifestyle of
T. gondii
has made them recalcitrant to molecular analysis in vivo.
Results
Here, we detail the results from transcriptional and proteomic analyses of bradyzoite-enriched fractions isolated from mouse brains infected with
T. gondii
over a time course of 21 to 150 days. The enrichment procedure afforded consistent identification of over 2000 parasitic peptides from the mixed-organism sample, representing 366
T. gondii
proteins at 28, 90, and 120 day timepoints. Deep sequencing of transcripts expressed during these three timepoints revealed that a subpopulation of genes that are transcriptionally expressed at a high level. Approximately one-third of these transcripts are more enriched during bradyzoite conditions compared to tachyzoites and approximately half are expressed at similar levels during each phase. The
T. gondii
transcript which increased the most over the course of chronic infection, sporoAMA1, shows stage specific isoform expression of the gene.
Conclusions
We have expanded the transcriptional profile of in vivo bradyzoites to 120 days post-infection and provided the first in vivo proteomic profile of
T. gondii
bradyzoites. The RNA sequencing depth of in vivo bradyzoite
T. gondii
was over 250-fold greater than previous reports and allowed us to identify low level transcripts and a novel bradyzoite-specific isoform of sporoAMA1.
Publisher
BioMed Central,BioMed Central Ltd,BMC
Subject
/ Animal Genetics and Genomics
/ Animals
/ Biomedical and Life Sciences
/ Computational Biology - methods
/ Cysts
/ Eukaryote microbial genomics
/ Gene Expression Profiling - methods
/ Genes
/ High-Throughput Nucleotide Sequencing
/ Life Cycle Stages - genetics
/ Mice
/ Microbial Genetics and Genomics
/ Neurons
/ Novels
/ Peptides
/ Proteins
/ Proteome
/ RNA
/ RNA-seq
/ Toxoplasma - growth & development
/ Toxoplasmosis - parasitology
/ Workflow
