Cyanobacteria, phototrophic organisms that perform oxygenic photosynthesis, perceive nitrogen status by sensing 2-oxoglutarate levels. PII, a widespread signaling protein, senses and transduces nitrogen and energy status to target proteins, regulating metabolism and gene expression. In cyanobacteria, under conditions of low 2-oxoglutarate, PII forms complexes with the enzyme N-acetyl glutamate kinase, increasing arginine biosynthesis, and with PII-interacting protein X (PipX), making PipX unavailable for binding and co-activation of the nitrogen regulator NtcA. Both the PII-PipX complex structure and in vivo functional data suggested that this complex, as such, could have regulatory functions in addition to PipX sequestration. To investigate this possibility we performed yeast three-hybrid screening of genomic libraries from Synechococcus elongatus PCC7942, searching for proteins interacting simultaneously with PII and PipX. The only prey clone found in the search expressed PlmA, a member of the GntR family of transcriptional regulators proven here by gel filtration to be homodimeric. Interactions analyses further confirmed the simultaneous requirement of PII and PipX, and showed that the PlmA contacts involve PipX elements exposed in the PII-PipX complex, specifically the C-terminal helices and one residue of the tudor-like body. In contrast, PII appears not to interact directly with PlmA, possibly being needed indirectly, to induce an extended conformation of the C-terminal helices of PipX and for modulating the surface polarity at the PII-PipX boundary, two elements that appear crucial for PlmA binding. Attempts to inactive plmA confirmed that this gene is essential in S. elongatus. Western blot assays revealed that S. elongatus PlmA, irrespective of the nitrogen regime, is a relatively abundant transcriptional regulator, suggesting the existence of a large PlmA regulon. In silico studies showed that PlmA is universally and exclusively found in cyanobacteria. Based on interaction data, on the relative amounts of the proteins involved in PII-PipX-PlmA complexes, determined in western assays, and on the restrictions imposed by the symmetries of trimeric PII and dimeric PlmA molecules, a structural and regulatory model for PlmA function is discussed in the context of the cyanobacterial nitrogen interaction network.
Las polifenol oxidasas (PPOs) son enzimas ubicuas que catalizan la reacción dependiente de oxígeno que transforma o-difenoles en o-quinonas. Estas quinonas son reactivas y capaces de modificar covalentemente un amplio abanico de especies nucleófilas, del interior de las células, que conduce a la formación de polímeros marrones, conocido como pardeamiento enzimático. El fenómeno de pardeamiento durante el crecimiento, recogida, almacenamiento y procesado de frutos y vegetales, es un problema de primera magnitud en la industria agroalimentaria y se reconoce como una de las principales causas de pérdidas de calidad y valor comercial. Produce cambios importantes tanto en la apariencia como en las propiedades organolépticas de frutos y vegetales comestibles, además suele ir asociado al desprendimiento de olores y efectos negativos sobre el valor nutricional. Aunque las PPOs se han descrito en diversos tejidos de plantas como raíces, semillas, hojas y frutos, el control de este fenómeno requiere un conocimiento bioquímico del tipo de sustratos fenólicos presentes en cada planta, el nivel de compuestos reductores, el nivel de accesibilidad del O2, de la naturaleza de los diferentes compuestos oxidables y de la polimerización y degradación de las o-quinonas. En este trabajo se presenta una revisión del efecto bioquímico, distribución, localización y posibles inhibidores de las PPOs en frutos y vegetales usados como alimento.
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