Enterohemorrhagic E. coli (EHEC) is an emerging zoonotic pathogen that causes bloody diarrhea and even death in humans worldwide. Currently, there is no targeted therapy and the use of antibiotics is constrained for EHEC infection, so understanding the pathogenesis of EHEC is required. Our previous studies have demonstrated that feeding C. elegans animals with EHEC causes ectopic expression of the microvillar actin (ACT-5) from the apical site to the cytoplasm in their intestinal cells. However, the host factors required for this EHEC-induced ACT-5 ectopic expression in C. elegans is still largely unknown. We therefore aimed to identify the host factors that are required for the EHEC-induced ACT-5 ectopic expression in C. elegans by genetic screening. We fed the fourth-stage C. elegans larvae with the mutagen, ethylmethane sulfonate (EMS), and then infected these animals with EHEC four days to select mutant worms confer normal ACT-5 expression. In several tests, we isolated two candidates, and named the mutations as the eae-1 (reduced ectopic-actin-expression-1) and eae-2 respectively. We first turned our attentions to the eae-1 allele, because it has significantly lower ratio of ACT-5 ectopic expression than wild type animals during EHEC infection. Interestingly, the total bacterial load in the eae-1 animals is not significant lower than that in the wild type animals after EHEC infection. However, the eae-1 animals are significantly resistant to EHEC infection. Taken all together, our current data suggested that the gene encoded by the eae-1 is most likely a specific host factor required for the EHEC-induced actin rearrangement in C. elegans.