RESUMEN
Emission and accumulation of carbon dioxide (CO2) in the atmosphere exert an environmental and climate change challenge. An attempt to deal with this challenge is made at Mongstad by application of amines for CO2 capture and storage (CO2 capture Mongstad (CCM) project). As part of the CO2 capture process, nitrosamines and nitramines may be emitted. Toxicological testing of nitrosamines and nitramines indicate a genotoxic potential of these substances. Here we present a risk characterization and assessment for five nitrosamines (N-Nitrosodi-methylamine (NDMA) N-Nitrosodi-ethylamine (NDEA), N-Nitroso-morpholine (NNM), N-Nitroso-piperidine (NPIP), and Dinitroso-piperazine (DNP)) and two nitramines (N-Methyl-nitramine (NTMA), Dimethyl-nitramine (NDTMA)), which are potentially emitted from the CO2 capture plant (CCP). Human health risk assessment of genotoxic non-threshold substances is a heavily debated topic, and no consensus methodology exists internationally. Extrapolation modeling from high-dose animal exposures to low-dose human exposures can be crucial for the final risk calculation. In the work presented here, different extrapolation models are discussed, and suggestions on applications are given. Then, preferred methods for calculating derived minimal effect level (DMEL) are presented with the selected nitrosamines and nitramines.
Asunto(s)
Compuestos de Anilina/toxicidad , Dióxido de Carbono/aislamiento & purificación , Nitrobencenos/toxicidad , Nitrosaminas/toxicidad , Compuestos de Anilina/administración & dosificación , Animales , Cambio Climático , Exposición a Riesgos Ambientales , Humanos , Pruebas de Mutagenicidad , Mutágenos/administración & dosificación , Mutágenos/toxicidad , Nitrobencenos/administración & dosificación , Nitrosaminas/administración & dosificación , Medición de RiesgoRESUMEN
Expression of the cobalamin (Cbl) biosynthetic cob operon in Salmonella typhimurium is repressed by the end-product. This regulation is conferred mainly at the translational level and involves a cobalamin-induced folding of an RNA hairpin that sequesters the ribosomal binding site (RBS) of the cob mRNA and prevents translation initiation. A combined structural and mutational analysis shows that a cis-acting translational enhancer (TE) element, located 83 nucleotides upstream of the Shine-Dalgarno sequence in the 5'-untranslated region (5'-UTR) of the cob mRNA, is required to unfold the inhibitory RBS hairpin in the absence of cobalamin. The TE element, which consists of 5 nucleotides, is proposed to confer its enhancer function in the absence of cobalamin by interacting with nucleotides in the stem of the RBS hairpin. This interaction destabilizes the RNA hairpin and allows ribosome binding. In the presence of cobalamin, the enhancer function is inhibited. As a result, the RBS hairpin forms and prevents translation initiation. Several additional RNA hairpins in the 5'-UTR were also identified and are suggested to be important for repression. The above data suggest that normal cobalamin repression of the cob operon requires that the 5'-UTR has a defined secondary and tertiary structure.
Asunto(s)
Apoproteínas/genética , Cobamidas/metabolismo , Grupo Citocromo b/genética , Elementos de Facilitación Genéticos , ARN Mensajero/química , Salmonella typhimurium/genética , Regiones no Traducidas 5' , Apoproteínas/metabolismo , Secuencia de Bases , Sitios de Unión , Grupo Citocromo b/metabolismo , Citocromos b , Datos de Secuencia Molecular , Mutación , Conformación de Ácido Nucleico , Biosíntesis de Proteínas , Ribosomas/metabolismoRESUMEN
Expression of the Salmonella typhimurium btuB gene, which encodes an outer membrane protein required for vitamin B12 uptake, is repressed by the presence of external vitamin B12. We have, by means of a mutational analysis, investigated which btuB sequences are required for repression. Analysis of btuB::lacZ transcriptional and translational fusions of various lengths showed that the control was exerted mainly at the translational level and required both coding and leader sequences in the btuB transcript. Regulatory mutants with a B12 non-repressible phenotype were isolated and the mutations were shown to be located at several sites within the btuB leader. Analysis of constructs carrying site-directed point mutations, which either destabilized or restabilized a putative RNA hairpin that sequesters the btuB ribosomal binding site, demonstrated that this hairpin was essential for normal repression. Comparison of the S. typhimurium btuB gene with the previously characterized S. typhimurium cbiA and Escherichia coli btuB genes reveals significant similarities as well as differences in the cis-acting sequences required for repression.
Asunto(s)
Proteínas de Escherichia coli , Biosíntesis de Proteínas , ARN Mensajero , Receptores de Péptidos/genética , Salmonella typhimurium/genética , Vitamina B 12/metabolismo , Proteínas de la Membrana Bacteriana Externa , Secuencia de Bases , Proteínas de Transporte de Membrana , Datos de Secuencia Molecular , Conformación de Ácido Nucleico , Mutación Puntual , ARN de Hongos , Salmonella typhimurium/metabolismoRESUMEN
Expression of the cob operon is repressed by B12 via a post-transcriptional control mechanism which requires sequence elements within the leader region of the mRNA and the first gene of the operon, the cbiA gene. Here we show that B12 repression of cbiA gene expression occurs at the level of translation initiation through sequestration of the ribosomal binding site (rbs) in an RNA hairpin. Analysis of mutations that destabilize or restabilize the secondary structure demonstrates that folding of the hairpin is essential for repression. The existence of the hairpin was confirmed by a secondary structure analysis of RNA from the wild type and three mutants. Deletions that remove the upstream part of the leader confer a drastic reduction in translation efficiency. This low-level translation is caused by the hairpin, as indicated by the finding that suppressor mutations that destabilize the hairpin restore efficient translation. Thus, the native upstream RNA functions as a translation enhancer and acts to relieve the hairpin's inhibitory effect on translation initiation. The inhibitory effect of the hairpin was confirmed by a ribosomal toeprinting analysis. We propose that the translational control of the cbiA gene mediates repression of the entire cob operon.