The Euonymus lectin (EUL) family is a unique group of carbohydrate-binding proteins that is omnipresent in plants. Sequences encoding EUL-related lectins have been retrieved from all completely sequenced plant genomes. The rice (Oryza sativa) genome contains 5 functional EUL genes referred to as OsEULS2, OsEULS3, OsEULD1a, OsEULD1b, and OsEULD2. In this study we focused on the tissue specific expression, stress inducibility and subcellular localization of the rice EULs. Even though the EUL domain sequence is highly conserved among the rice EULs (at least 80% sequence similarity) different biotic and abiotic stress treatments yielded unique responses for the different EULs. Transcript levels for OsEULs were differentially affected by drought and salt stress, ABA treatment, pathogen infection or insect infestation. Analysis of promoter activity revealed differential expression and tissue specificity for the 5 OsEUL genes, with most expression observed in the vascular system of roots and shoots, as well as in the root tips and seeds. At cell level, all OsEULs are located in the nucleus whereas OsEULD1b and OsEULD2 also locate to the cytoplasm. This paper contributes to the functional characterization of the EULs and provides insight in the biological importance of this family of proteins for rice.

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Ribosome-inactivating proteins (RIPs) are a class of cytotoxic enzymes that can depurinate rRNAs thereby inhibiting protein translation. Although these proteins have also been detected in bacteria, fungi, and even some insects, they are especially prevalent in the plant kingdom. This review focuses on the RIPs from cereals. Studies on the taxonomical distribution and evolution of plant RIPs suggest that cereal RIPs have evolved at an enhanced rate giving rise to a large and heterogeneous RIP gene family. Furthermore, several cereal RIP genes are characterized by a unique domain architecture and the lack of a signal peptide. This advanced evolution of cereal RIPs translates into distinct structures, activation mechanisms, and physiological roles. Several cereal RIPs are characterized by activation mechanisms that include the proteolytic removal of internal peptides from the N-glycosidase domain, a feature not documented for non-cereal RIPs. Besides their role in defense against pathogenic fungi or herbivorous insects, cereal RIPs are also involved in endogenous functions such as adaptation to abiotic stress, storage, induction of senescence, and reprogramming of the translational machinery. The unique properties of cereal RIPs are discussed in this review paper.

Ribosome-inactivating proteins (RIPs) are cytotoxic enzymes that inhibit protein translation by depurinating ribosomal RNA. Although most plant RIPs are synthesized with leader sequences that sequester them away from the host ribosomes, several RIPs from cereals lack these signal peptides and therefore probably reside in the cytosol near the plant ribosomes. More than 30 RIP genes have been identified in the rice (Oryza sativa spp. japonica) genome, many of them lacking a signal peptide. This paper focuses on a presumed cytosolic type-1 RIP from rice, referred to as OsRIP1. Using 3D modeling it is shown that OsRIP1 structurally resembles other cereal RIPs and has an active site that meets the requirements for activity. Furthermore, localization studies indicate that OsRIP1-eGFP fusion proteins reside in the nucleocytoplasmic space when expressed in epidermal cells of Nicotiana benthamiana or Arabidopsis thaliana suspension cells. Finally, OsRIP1 was recombinantly produced in Escherichia coli and was demonstrated to possess catalytic activity. Interestingly, this recombinant RIP inactivates wheat ribosomes far less efficiently than rabbit ribosomes in an in vitro system. These findings raise some interesting questions concerning the mode of action and physiological role of OsRIP1. This is the first time a RIP from rice is investigated at protein level and is shown to possess biological activity.

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