Identification and in silico Analysis of NADPH Oxidase Homologues Involved in Allergy from an Olive Pollen Transcriptome

Abstract

Abstract. Reactive oxygen species generated by pollen NADPH oxidases are present in numerous allergenic pollen species. The superoxide generated by this enzyme has been suggested as a key actor in the induction of allergic inflammation. However, this enzyme has been characterized in Arabidopsis thaliana pollen only, where two pollen-specific genes (RbohH and RbohJ) have been described. The olive (Olea europaea L.) pollen is an important source of allergy in Mediterranean countries. We have assembled and annotated an olive pollen transcriptome, which allowed us to determine the presence of at least two pollen-specific NADPH oxidase homologues. Primers were designed to distinguish between the two homologues, and full-length sequences were obtained through a PCR strategy. Complete in silico analysis of such sequences, including phylogeny, 3-D modeling of the N-terminus, and prediction of cellular localization and post-translational modifications was carried out with the purpose of shed light into the involvement of olive pollen-intrinsic NADPH oxidases in triggering allergy symptoms.