Characterization of a secretory type Theileria parva glutaredoxin homologue identified by novel screening procedure.
The schizont stage of the protozoan parasite Theileria parva induces features characteristic of tumor cells in infected bovine T-cell lines. Most strikingly T. parva-infected cell lines acquire unlimited growth potential in vitro. Their proliferative state is entirely dependent on the presence of a viable parasite within the host cell cytoplasm. It has been postulated that parasite proteins either secreted into the host cell or expressed on the parasite surface membrane are involved in the parasite-host cell interaction. We used an in vitro transcription-translation-membrane translocation system to identify T. parva-derived cDNA clones encoding secretory or membrane proteins. Within 600 clones we found one encoding a 17-kDa protein which is processed by microsomal membranes to a 14-kDa protein (11E), presumably by signal peptidase. The processed form is expressed in the T-cell line TpM803 harboring viable parasites. By immunolocalization we show that the 11E protein mostly resides within the parasite, often in close vicinity to membranous structures, but in addition it appears at the surface membrane. Amino acid sequence comparison suggests that 11E belongs to the glutaredoxin family, but is unique so far in containing a signal sequence for endoplasmic reticulum membrane translocation.
Ebel
T
T
Vienna International Research Cooperation Center, University of Vienna, A-1235 Vienna, Austria.
Middleton
J F
JF
Frisch
A
A
Lipp
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000032:analysis / Q000096:biosynthesis; D002417:Cattle; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D003001:Cloning, Molecular; D016384:Consensus Sequence; D015723:Gene Library; D054477:Glutaredoxins; D007425:Intracellular Membranes / Q000378:metabolism; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000737:chemistry; D008861:Microsomes / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010088:Oxidoreductases / Q000737:chemistry; D014176:Protein Biosynthesis; D011490:Protein Disulfide Reductase (Glutathione); D011506:Proteins / Q000737:chemistry*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D013601:T-Lymphocytes; D016797:Theileria parva / Q000378:metabolism*; D014158:Transcription, Genetic
D000953:Antigens, Protozoan; D054477:Glutaredoxins; D008565:Membrane Proteins; D011506:Proteins; D011994:Recombinant Proteins; D010088:Oxidoreductases; D011490:Protein Disulfide Reductase (Glutathione)