Differential immune-reactivity and subcellular distribution reveal the multifunctional character of profilin in pollen as major effect of sequences polymorphism

Abstract

Profilin, one of the major allergen (Ole e 2) of olive (Olea europaea L.) pollen, are broadly distributed actin-monomer-binding proteins (ABP). They display a major regulatory role in actin cytoskeleton dynamics, driving cell morphogenesis, sexual reproduction, and translating signals into cellular responses to different environmental stresses. Plants exhibit multiple profilin isoforms w ith distinctive biochemical properties, and differentially regulated. How ever, it is still an open question w hether these profilin isoforms, generated by multiple gene sequence polymorphism, are functionally different, as well as the role of that polymorphism in pollen allergy. Particularly, in differential epitopes generation, profilin isoforms sensitization and cross-reactivity among cultivars, and even among species. In the present study, w e have used mature pollen from olive, birch, hazel, timothy-grass, and maize, in addition to olive germinating pollen and seeds, w ith the aim to analyze the immune-reactivity and subcellular localization of profilin by using polyclonal and specific isoforms antibodies against olive and maize profilins. The results show ed immune-reactivity differences betw een the five species analyzed, betw een olive cultivars, as w ell as between reproductive and vegetative profilins. Furthermore, the existence of different profilin isoforms w as revealed along pollen germination stages. A differential subcellular distribution of profilin isoforms w as found in olive pollen. They w ere localized in the nucleus, pollen aperture regions, pollen and tube w alls and pollen tip, in addition to a general cytoplasmic distribution, in comparison to controls. Data suggest that profilin family might contain numerous functionally distinctive isoforms, spatial-temporal differentially expressed and regulated during vegetative development, pollen maturation and pollen tube grow th. Furthermore, differential immune-reactivity revealed in the study might point out the involvement of common shared and specific epitopes, generated by sequence polymorphism, in differential olive pollen cultivar sensitization of allergenic patients, and cross-reaction to pollen from different species.