A Giardia duodenalis gene encoding a protein with multiple repeats of a toxin homologue

A gene, CRP136, from the intestinal protozoan parasite Giardia duodenalis, expressed at a high level in a metronidazole-resistant line, encodes a 136 kDa protein with 23 copies of a 40 amino acid repeat. The protein is cysteine-rich and has the typical membrane-spanning region and CXXC amino acid motifs of a family of Giardia cysteine-rich surface proteins (CRSPs). The repeat unit in CRP136, shares 57% homology with the gene encoding the precursor of the sarafotoxins, a group of snake toxins from the burrowing adder known to cause symptoms similar to those of humans acutely infected with Giardia. The sarafotoxins are low molecular weight sulphydryl cross-linked peptides which are proteolytically cleaved from a precursor polyprotein. CRP136 has homology over the entire length of the sarafotoxin precursor, and the repeats are of the same length. Thus CRP136 represents the first evidence for a potential Giardia toxin. The genomic copy number of CRP136 appears to be the same in both the parent and drug-resistant lines and expression of this gene, and at least one other, is associated with a conserved partial duplication, but not amplification, of one chromosome.