RESUMEN
NUDEL-oligopeptidase is a cytosolic cysteine peptidase, active towards oligopeptides and involved in the conversion and inactivation of a number of bioactive peptides. This protein interacts with neuronal proteins and is essential for brain development and cortical organization during embryogenesis. In this study, 5'-flanking sequences of the human and rabbit NUDEL-oligopeptidase gene were cloned into the pGL3 reporter gene vector and the promoter activity of the full-length fragment and deletions series was measured in transient transfection assays using two different cell lines, namely, C6 rat glioma and NH15 human neuroblastoma. Overall, a very similar pattern of promoter activity was obtained for both rabbit and human NUDEL-oligopeptidase promoter sequences, and their respective serial deletion constructs upon transient transfection into these cell lines. The only exception was for the longest rabbit upstream sequence that displayed about 1.8-fold higher luciferase expression upon transfection into NH15 neuronal cells than that observed upon transfection into C6 glioma cells. On the other hand, no significant difference was observed for the human longest sequence. These results are in good agreement with the expression pattern of NUDEL-oligopeptidase in human and rabbit tissues.
Asunto(s)
Proteínas Portadoras/genética , Regulación de la Expresión Génica , Regiones Promotoras Genéticas/genética , Animales , Secuencia de Bases , Northern Blotting , Línea Celular Tumoral , Sistema Nervioso Central/metabolismo , Clonación Molecular , Vectores Genéticos/genética , Humanos , Luciferasas , Datos de Secuencia Molecular , Conejos , Ratas , Análisis de Secuencia de ADNRESUMEN
OBJECTIVES: There is controversial evidence regarding the levels of antioxidant molecules in type 2 diabetes periodontitis patients. Thus, the aim of the present study was to evaluate the gene expression of antioxidant enzymes in the gingival tissue of poorly and well-controlled type 2 diabetic subjects with chronic periodontitis (CP). DESIGN: Gingival biopsies were harvested from systemically and periodontally healthy subjects (n=12), systemically healthy subjects with CP (n=15), well-controlled (n=8) and poorly controlled (n=14) diabetic subjects with CP. The messenger RNA (mRNA) levels of peroxiredoxin (PRDX) 1 and 2, catalase (CAT), glutathione peroxidase (GPX1) and superoxide dismutase (SOD) 1 and 2 were measured by quantitative polymerase chain reaction (qPCR). RESULTS: The results showed that PRDX1 and GPX1 were up-regulated by periodontitis (p<0.05), independently of the glycaemic status, whilst PRDX2 and SOD2 genes were slightly influenced by periodontitis, but significantly induced when periodontitis was associated with DM, especially under a poor glycaemic control (p<0.05). Moreover, CAT and SOD1 expressions were not significantly influenced by any of these inflammatory disorders (p>0.05). CONCLUSION: In conclusion, both PRDX1 and GPX1 were overexpressed in CP whilst PRDX2 and SOD2 were up-regulated especially in the poorly controlled diabetic group with CP.
Asunto(s)
Antioxidantes/metabolismo , Periodontitis Crónica/enzimología , Diabetes Mellitus Tipo 2/enzimología , Encía/enzimología , Adulto , Biopsia , Estudios de Casos y Controles , Catalasa/genética , Catalasa/metabolismo , Periodontitis Crónica/complicaciones , Diabetes Mellitus Tipo 2/complicaciones , Femenino , Expresión Génica , Glutatión Peroxidasa/genética , Glutatión Peroxidasa/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Estrés Oxidativo , Peroxirredoxinas/genética , Peroxirredoxinas/metabolismo , Reacción en Cadena de la Polimerasa , ARN Mensajero/metabolismo , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismoRESUMEN
Recently, nuclear distribution element-like (NUDEL) has been implicated to play a role in lissencephaly and schizophrenia through interactions with the lissencephaly gene 1 (Lis1) and disrupted-in-schizophrenia 1 (DISC1) products, respectively. Interestingly, NUDEL is the same protein as endooligopeptidase A (EOPA), a thiol-activated peptidase involved in conversion and inactivation of a number of bioactive peptides. In this study, we have cloned EOPA from the human brain and have confirmed that it is equivalent to NUDEL, leading us to suggest a single name, NUDEL-oligopeptidase. In the brain, the monomeric form of NUDEL-oligopeptidase is responsible for the peptidase activity whose catalytic mechanism is likely to involve a reactive cysteine, because mutation of Cys-273 fully abolished NUDEL-oligopeptidase activity without disrupting the protein's secondary structure. Cys-273 is very close to the DISC1-binding site on NUDEL-oligopeptidase. Intriguingly, DISC1 inhibits NUDEL-oligopeptidase activity in a competitive fashion. We suggest that the activity of NUDEL-oligopeptidase is under tight regulation through protein-protein interactions and that disruption of these interactions, as postulated in a Scottish DISC1 translocation schizophrenia cohort, may lead to aberrant regulation of NUDEL-oligopeptidase, perhaps providing a substrate for the pathology of schizophrenia.