ABSTRACT
New single-chain (type 1) ribosome-inactivating proteins (RIPs) were isolated from the seeds of Basella rubra L. (two proteins) and from the leaves of Bougainvillea spectabilis Willd. (one protein). These RIPs inhibit protein synthesis both in a cell-free system, with an IC50 (concentration causing 50% inhibition) in the 10(-10) M range, and by various cell lines, with IC50S in the 10(-8)-10(-6) M range. All three RIPs released adenine not only from rat liver ribosomes but also from Escherichia coli rRNA, polyadenylic acid, herring sperm DNA, and artichoke mottled crinkle virus (AMCV) genomic RNA, thus being polynucleotide:adenosine glycosidases. The proteins from Basella rubra had toxicity to mice similar to that of most type 1 RIPs (Barbieri et al., 1993, Biochim Biophys Acta 1154: 237-282) with an LD50 (concentration that is 50% lethal) < or = 8 mg.kg-1 body weight, whilst the RIP from Bougainvillea spectabilis had an LD50 > 32 mg.kg-1. The N-terminal sequence of the two RIPs from Basella rubra had 80-93% identity, whereas it differed from the sequence of the RIP from Bougainvillea spectabilis. When tested with antibodies against various RIPs, the RIPs from Basella gave some cross-reactivity with sera against dianthin 32, and weak cross-reactivity with momordin I and momorcochin-S, whilst the RIP from Bougainvillea did not cross-react with any antiserum tested. An RIP from Basella rubra and one from Bougainvillea spectabilis were tested for antiviral activity, and both inhibited infection of Nicotiana benthamiana by AMCV.
Subject(s)
Antiviral Agents/pharmacology , N-Glycosyl Hydrolases/metabolism , Plants/enzymology , Protein Synthesis Inhibitors/pharmacology , 3T3 Cells , Amino Acid Sequence , Animals , Antiviral Agents/chemistry , Antiviral Agents/isolation & purification , DNA/metabolism , Female , HeLa Cells , Humans , Male , Mice , Molecular Sequence Data , N-Glycosyl Hydrolases/chemistry , N-Glycosyl Hydrolases/isolation & purification , N-Glycosyl Hydrolases/pharmacology , Plant Proteins/chemistry , Plant Proteins/isolation & purification , Plant Proteins/pharmacology , Poly A/metabolism , Protein Synthesis Inhibitors/chemistry , Protein Synthesis Inhibitors/isolation & purification , RNA, Bacterial/metabolism , RNA, Viral/metabolism , Rabbits , Rats , Ribosome Inactivating Proteins , Ribosomes , Tumor Cells, CulturedABSTRACT
The phenotypic characteristics of four Burkholderia cepacia strains isolated from the rhizosphere and the clinical environment were compared. Tests included optimum growth temperature, utilization of carbon sources, production of HCN, indole-3-acetic acid (IAA) and siderophores, proteolytic activity, nitrogen fixation, inhibition of some phytopathogenic fungi, adherence to human mucosal and plant root epithelia, and greenhouse-based plant-growth promotion experiments using cucumber (Cucumis sativus). Results indicated that the strains of B. cepacia isolated from the rhizosphere differ markedly from their clinical counterparts. Strains isolated from the rhizosphere grew over a wider temperature range, fixed nitrogen and produced IAA, did not produce proteases, displayed a wider antibiosis against the phytopathogenic fungi studied, did not adhere to human uroepithelial cells, promoted growth of C. sativus and only produced a hydroxamate-like siderophore. In contrast, clinical isolates could not fix nitrogen or produce IAA, produced proteases, adhered to human uroepithelial cells, did not promote the growth of C. sativus and, in addition to a hydroxamate-like siderophore, produced pyochelin and salicylate siderophores. All four isolates exhibited the ability to adhere to the root tissue of C. sativus and were unable to produce HCN.
