Silencing of GRA10 protein expression inhibits Toxoplasma gondii intracellular growth and development
| Autor: | Kwame Matthews, Mark McHugh, William H. Witola, Bretta Bauman |
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| Rok vydání: | 2014 |
| Předmět: |
Male
Molecular Sequence Data Protozoan Proteins Antibodies Protozoan Biology Monocytes parasitic diseases Animals Humans Gene silencing Amino Acid Sequence Gene Silencing Cloning Molecular Cells Cultured Messenger RNA Gene knockdown Sheep Toxoplasma gondii Fibroblasts biology.organism_classification Molecular biology Rats Cell biology Cytosol Infectious Diseases Polyclonal antibodies biology.protein Parasitology Rabbits Dense granule Toxoplasma Intracellular |
| Zdroj: | Parasitology International. 63:651-658 |
| ISSN: | 1383-5769 |
| DOI: | 10.1016/j.parint.2014.05.001 |
| Popis: | Toxoplasma gondii dense granule proteins (GRAs) are secreted abundantly in both the tachyzoite and bradyzoite stages of the parasite and are known to localize to various compartments of the parasitophorous vacuole (PV) that interfaces with the host cell milieu. Thus, GRAs may play significant roles in the biogenesis of the PV that is important for survival of intracellular T. gondii. GRA10 is a dense granule protein whose role in T. gondii has not yet been characterized. Therefore, in this study, we endeavored to determine the role of GRA10 in the growth and survival of intracellular T. gondii by using phosphorodiamidate morpholino oligomers (PPMOs) antisense knockdown approach to disrupt the translation of GRA10 mRNA in the parasites. We expressed and purified a truncated recombinant GRA10 protein to generate anti-GRA10 polyclonal antibodies that we used to characterize GRA10 in T. gondii. We found that GRA10 is a soluble, dense granule-associated protein that is secreted into the parasite cytosol and the parasitophorous vacuole milieu. Using in vitro cultures, we found that knockdown of GRA10 results in severe inhibition of T. gondii growth in human fibroblasts and in ovine monocytic cells. Together, our findings define GRA10 as a dense granule protein that plays a significant role in the growth and propagation of intracellular T. gondii in human fibroblasts and in ovine monocytic cells. |
| Databáze: | OpenAIRE |
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