Eukaryot Cell. 2011 Oct 21. [Epub ahead of print]
Identification of tissue cyst wall components by transcriptome analysis of in vivo and in vitro Toxoplasma bradyzoites.
Buchholz KR, Fritz HM, Chen X, Durbin-Johnson B, Rocke DM, Ferguson DJ, Conrad PA, Boothroyd JC.
SourceDepartment of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, 94305, USA.
The Toxoplasma gondii bradyzoite is essential to establish persistent infection yet little is known about what factors this developmental form secretes to establish the cyst or interact with its host cell. To identify candidate bradyzoite-secreted effectors, the transcriptomes of in vitro tachyzoites 2 days post-infection, in vitro bradyzoites 4 days post-infection, and in vivo bradyzoites 21 days post-infection were interrogated by microarray, and the program SignalP was used to identify signal peptides indicating secretion. One-hundred-and-two putative bradyzoite-secreted effectors were identified with this approach. Two candidates, bradyzoite pseudokinase 1 (BPK1) and microneme adhesive repeat (MAR)-domain containing protein 4 (MCP4), were chosen for further investigation and confirmed to be induced and secreted by bradyzoites in vitro and in vivo. Thus, we report the first analysis of the transcriptomes of in vitro and in vivo bradyzoites and identify two new protein components of the Toxoplasma tissue cyst wall.
PMID:22021236[PubMed - as supplied by publisher]