Ribosome-inactivating proteins (RIPs) are a large group of plant enzymes present in a great variety of species, which inhibit protein synthesis through a site-specific deadenylation of the large ribosomal RNA at level of the conserved alpha-sarcin/ricin loop. Saporins are highly basic single chain RIPs present in different organs of the plant Saponaria officinalis (Caryiophyllacae). Previously in our laboratory, two leaf isoforms, saporin L3 and saporin S6-like, characterized by different toxicity profiles were isolated. These isoforms were isolated from intra and extracellular fractions, respectively. We constructed GFP fusions to obtain more detailed information on the localization of these two saporins. Transformation of Arabidopsis thaliana seedling roots using Agrobacterium and of onion epidermal tissues using biolistics were performed. In roots, our preliminary data on the fusions with the NH-terminus portion of two saporins (first 100 aa of the immature protein with signal peptide) suggests this region is sufficient for intra- and extracellular targeting. A potential tissue dependent targeting of saporin S6-like derived from onion epidermal data is presented.
LEAF SAPORIN ISOFORMS: CELLULAR LOCALIZATION BY GFP-FUSION ANALYSIS.
RODRIGUES POUSADA, RENATO ALBERTO;
2010-01-01
Abstract
Ribosome-inactivating proteins (RIPs) are a large group of plant enzymes present in a great variety of species, which inhibit protein synthesis through a site-specific deadenylation of the large ribosomal RNA at level of the conserved alpha-sarcin/ricin loop. Saporins are highly basic single chain RIPs present in different organs of the plant Saponaria officinalis (Caryiophyllacae). Previously in our laboratory, two leaf isoforms, saporin L3 and saporin S6-like, characterized by different toxicity profiles were isolated. These isoforms were isolated from intra and extracellular fractions, respectively. We constructed GFP fusions to obtain more detailed information on the localization of these two saporins. Transformation of Arabidopsis thaliana seedling roots using Agrobacterium and of onion epidermal tissues using biolistics were performed. In roots, our preliminary data on the fusions with the NH-terminus portion of two saporins (first 100 aa of the immature protein with signal peptide) suggests this region is sufficient for intra- and extracellular targeting. A potential tissue dependent targeting of saporin S6-like derived from onion epidermal data is presented.Pubblicazioni consigliate
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