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Background  Cryptosporidiosis is a ubiquitous infectious disease, caused by the protozoan parasites  Cryptosporidium hominis  and  C .  parvum , leading to acute, persistent and chronic diarrhea worldwide. Although the complications of this disease can be serious, even fatal, in immunocompromised patients of any age, they have also been found to lead to long term effects, including growth inhibition and impaired cognitive development, in infected immunocompetent children. The  Cryptosporidium  life cycle alternates between a dormant stage, the oocyst, and a highly replicative phase that includes both asexual vegetative stages as well as sexual stages, implying fine genetic regulatory mechanisms. The parasite is extremely difficult to study because it cannot be cultured  in vitro  and animal models are equally challenging. The recent publication of the genome sequence of  C. hominis  and  C. parvum  has, however, significantly advanced our understanding of the biology and pathogenesis of this parasite.    Methodology/Principal Findings  Herein, our goal was to identify  cis -regulatory elements associated with heat shock response in  Cryptosporidium  using a combination of  in silico  and real time RT-PCR strategies. Analysis with Gibbs-Sampling algorithms of upstream non-translated regions of twelve genes annotated as heat shock proteins in the  Cryptosporidium  genome identified a highly conserved over-represented sequence motif in eleven of them. RT-PCR analyses, described herein and also by others, show that these eleven genes bearing the putative element are induced concurrent with excystation of parasite oocysts via heat shock.    Conclusions/Significance  Our analyses suggest that occurrences of a motif identified in the upstream regions of the  Cryptosporidium  heat shock genes represent parts of the transcriptional apparatus and function as stress response elements that activate expression of these genes during excystation, and possibly at other stages in the life cycle of the parasite. Since heat shock and excystation represent a critical step in the development of the infectious sporozoite form of  Cryptosporidium , these results provide important insight into the pathogenicity of the parasite.

Putative cis-Regulatory Elements Associated with Heat Shock Genes Activated During Excystation of Cryptosporidium parvum