Comparative and evolutionary analysis of Arabidopsis RIN4-like/NOI proteins induced by herbivory.

The spider mite Tetranychus urticae is an economically important agricultural pest, which feeds on a broad spectrum of plant species. In an RNAseq experiment performed in our laboratory, 4 of the 15 members of the RIN4-like/NOI family of Arabidopsis thaliana were significantly overexpressed after T. urticae infestation. Two of them (NOI3 and NOI5) are shorter and harbour one NOI domain, which characterises this family, and the other two (NOI10 and NOI11) have two-NOI domains. The only member of this family characterized is RIN4, a two-NOI intrinsically disordered protein anchored to the plasma membrane and involved in plant defence against bacterial pathogens. The function of all other members of the RIN4-like/NOI Arabidopsis family and their putative role in herbivore defence remains unknown. We perform a comparative genomic analysis of RIN4-like/NOI sequences to study the evolutionary features of this protein family and the distribution of its members among species. We show that short one-NOI proteins were more numerous and exhibited lower disorder propensity compared to two-NOI members. NOI10 and NOI11, from the two-NOI group, are included in a clade-specific expansion of Brassicaceae with unique predicted posttranslational modification sites and clear predicted structural differences from RIN4. Our analysis suggests that the members of the RIN4-like/NOI family upregulated after mite feeding have novel functions different from those assigned to RIN4, likely involving adaptation to stress specialisation.