Functional interactions between members of the REPAT family of insect pathogen-induced proteins.

Studies on the transcriptional response to pathogens in the insect larval gut have shown the regulation of several genes after the infection. Repat (REsponse to PAThogens) genes were first identified in Spodoptera exigua midgut as being up-regulated in response to the exposure to Bacillus thuringiensis toxins and baculovirus. Recently, new members of the REPAT family showed a constitutive up-regulation in a B. thuringiensis-resistant population. Based on a yeast two-hybrid screening, we have detected the interaction of REPAT1 with other members of the REPAT family, leading to the discovery of a new member: REPAT8. The functional role of this interaction was shown by following the changes of the subcellular localization of REPAT1 in the presence of REPAT8. REPAT1 alone was localized exclusively in the cytoplasm, while the presence of REPAT8 led to the migration of REPAT1 to the nucleus. Finally, analysis of the expression pattern of eight REPAT members has shown that B. thuringiensis-related treatments (Cry1Ca toxin, Xentari™ product and an acrystalliferous strain) induced a general up-regulation of repat genes, especially of repat2. In contrast, no significant effect was detected after treatment with Escherichia coli or Enterococcus sp., or by the presence of microbiota in the midgut. The results suggest that the different repat genes play different roles in response to pathogens.