Expression and characterization of a novel teratocyte protein of the braconid, Microplitis croceipes (cresson).
Microplitis croceipes wasps overcome host immunity by inducing changes in host physiology using factors derived from the embryo and/or larva. Teratocytes of some parasitic wasps circulate in the host hemolymph after egg hatch and synthesize proteins (TSPs), some of which are secreted to alter host physiology in support of endoparasitoid development. TSPs appear to alter host physiology, at least in part, by inhibiting synthesis of certain proteins. M. croceipes teratocytes synthesize a 13.9 kDa protein (TSP14), which inhibits synthesis of host proteins that are linked to larval growth and development. A cDNA encoding TSP14 was generated by RT-PCR from teratocyte RNA, and cloned into yeast expression vectors to produce sufficient recombinant protein for functional analyses. RecTSP14 was produced using the yeast expression system at a concentration of more than 300 micrograms/L. The recTSP14 inhibited in vitro translation of larval Heliothis virescens RNA, with the activity sensitive to boiling, protein concentration, incubation time, and storage temperatures. Although recTSP14 inhibited translation of some cellular RNAs in vitro, the in vivo incorporation of [35S]-methionine into proteins of selected insect and mammalian cell lines was not inhibited. These findings suggest that recTSP14 entry is cell type-specific and required to inhibit synthesis of target protein(s).
Rana
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RL
Department of Entomology, University of Kentucky, Lexington, KY 40546-0091, USA. rlrana2@uky.edu
Dahlman
D L
DL
Webb
B A
BA
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D006458:Hemolymph / Q000502:physiology; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism*; D008715:Methionine / Q000378:metabolism; D010058:Oviposition; D014176:Protein Biosynthesis; D014162:Transfection; D014407:Tumor Cells, Cultured; D014863:Wasps / Q000235:genetics / Q000378:metabolism*
D017931:DNA Primers; D018076:DNA, Complementary; D019476:Insect Proteins; D008715:Methionine