Identification of novel bradyzoite-specific Toxoplasma gondii genes with domains for protein–protein interactions by suppression subtractive hybridization

Abstract

By using suppression subtractive hybridization we identified five so far uncharacterized stage specific genes in Toxoplasma gondii, which are induced during tachyzoite-to-bradyzoite differentiation. The mRNA level of a putative zinc-finger protein was increased 23-fold in bradyzoites, while the remaining four genes displayed induction levels >100-fold. Two of these genes predict proteins with domains for protein–protein interactions. One protein (ANK1) contains both, a TPR-domain and an ankyrin motif, which consists of seven repeats. ANK1 was shown by epitope tagging experiments to be localized in the cytosol. In a fraction of parasites, the myc-tagged fusion protein was additionally localized in the nucleus, which is in agreement with the presence of a bipartite nuclear targeting sequence in ANK1. The identification of bradyzoite-specific proteins with TPR- and ankyrin-domains supports the concept that during stage conversion a variety of proteins which are involved in protein–protein interactions are induced, thereby assisting the rebuilding of the proteome.