RESUMEN
Sister chromatid cohesion (SCC), efficient DNA repair, and the regulation of some metazoan genes require the association of cohesins with chromosomes. Cohesins are deposited by a conserved heterodimeric loading complex composed of the Scc2 and Scc4 proteins in Saccharomyces cerevisiae, but how the Scc2/Scc4 deposition complex regulates the spatiotemporal association of cohesin with chromosomes is not understood. We examined Scc2 chromatin association during the cell division cycle and found that the affinity of Scc2 for chromatin increases biphasically during the cell cycle, increasing first transiently in late G1 phase and then again later in G2/M. Inactivation of Scc2 following DNA replication reduces cellular viability, suggesting that this post S-phase increase in Scc2 chromatin binding affinity is biologically relevant. Interestingly, high and low Scc2 chromatin binding levels correlate strongly with the presence of full-length or amino-terminally cleaved forms of Scc2, respectively, and the appearance of the cleaved Scc2 species is promoted in vitro either by treatment with specific cell cycle-staged cellular extracts or by dephosphorylation. Importantly, Scc2 cleavage eliminates Scc2-Scc4 physical interactions, and an scc2 truncation mutant that mimics in vivo Scc2 cleavage is defective for cohesin deposition. These observations suggest a previously unidentified mechanism for the spatiotemporal regulation of cohesin association with chromosomes through cell cycle regulation of Scc2 cohesin deposition activity by Scc2 dephosphorylation and cleavage.
Asunto(s)
Proteínas de Ciclo Celular/metabolismo , Proteínas Cromosómicas no Histona/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/metabolismo , Secuencia de Aminoácidos , Animales , Ciclo Celular/fisiología , Cromatina/metabolismo , Inestabilidad Cromosómica/fisiología , Proteínas Cromosómicas no Histona/genética , Segregación Cromosómica/fisiología , Genotipo , Datos de Secuencia Molecular , Fosforilación/fisiología , Conejos , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/genética , CohesinasRESUMEN
Parkinson disease is caused by the death of midbrain dopamine neurons from oxidative stress, abnormal protein aggregation, and genetic predisposition. In 2003, Bonifati et al. (23) found that a single amino acid mutation in the DJ-1 protein was associated with early-onset, autosomal recessive Parkinson disease (PARK7). The mutation L166P prevents dimerization that is essential for the antioxidant and gene regulatory activity of the DJ-1 protein. Because low levels of DJ-1 cause Parkinson, we reasoned that overexpression might stop the disease. We found that overexpression of DJ-1 improved tolerance to oxidative stress by selectively up-regulating the rate-limiting step in glutathione synthesis. When we imposed a different metabolic insult, A53T mutant α-synuclein, we found that DJ-1 turned on production of the chaperone protein Hsp-70 without affecting glutathione synthesis. After screening a number of small molecules, we have found that the histone deacetylase inhibitor phenylbutyrate increases DJ-1 expression by 300% in the N27 dopamine cell line and rescues cells from oxidative stress and mutant α-synuclein toxicity. In mice, phenylbutyrate treatment leads to a 260% increase in brain DJ-1 levels and protects dopamine neurons against 1-methyl 4-phenyl 1,2,3,6-tetrahydropyridine (MPTP) toxicity. In a transgenic mouse model of diffuse Lewy body disease, long-term administration of phenylbutyrate reduces α-synuclein aggregation in brain and prevents age-related deterioration in motor and cognitive function. We conclude that drugs that up-regulate DJ-1 gene expression may slow the progression of Parkinson disease by moderating oxidative stress and protein aggregation.
Asunto(s)
Proteínas Oncogénicas/efectos de los fármacos , Enfermedad de Parkinson/tratamiento farmacológico , Fenilbutiratos/farmacología , Animales , Células Cultivadas , Inhibidores de Histona Desacetilasas , Ratones , Neuronas , Fármacos Neuroprotectores , Proteínas Oncogénicas/biosíntesis , Estrés Oxidativo/efectos de los fármacos , Peroxirredoxinas , Fenilbutiratos/uso terapéutico , Proteína Desglicasa DJ-1 , Proteínas/metabolismo , Regulación hacia Arriba/efectos de los fármacosRESUMEN
Reducing insulin/IGF signaling allows for organismal survival during periods of inhospitable conditions by regulating the diapause state, whereby the organism stockpiles lipids, reduces fertility, increases stress resistance, and has an increased lifespan. The Target of Rapamycin (TOR) responds to changes in growth factors, amino acids, oxygen tension, and energy status; however, it is unclear how TOR contributes to physiological homeostasis and disease conditions. Here, we show that reducing the function of Drosophila TOR results in decreased lipid stores and glucose levels. Importantly, this reduction of dTOR activity blocks the insulin resistance and metabolic syndrome phenotypes associated with increased activity of the insulin responsive transcription factor, dFOXO. Reduction in dTOR function also protects against age-dependent decline in heart function and increases longevity. Thus, the regulation of dTOR activity may be an ancient "systems biological" means of regulating metabolism and senescence, that has important evolutionary, physiological, and clinical implications.
Asunto(s)
Proteínas de Drosophila/metabolismo , Factores de Transcripción Forkhead/metabolismo , Resistencia a la Insulina/fisiología , Fosfatidilinositol 3-Quinasas/metabolismo , Alelos , Secuencia de Aminoácidos , Animales , Regulación hacia Abajo , Drosophila , Proteínas de Drosophila/antagonistas & inhibidores , Proteínas de Drosophila/genética , Proteínas de Drosophila/farmacología , Factores de Transcripción Forkhead/antagonistas & inhibidores , Glucosa/análisis , Lípidos/análisis , Modelos Biológicos , Datos de Secuencia Molecular , Fosfatidilinositol 3-Quinasas/genética , Fosfatidilinositol 3-Quinasas/farmacología , Proteínas Quinasas , Alineación de Secuencia , Transducción de Señal , Serina-Treonina Quinasas TOR , Regulación hacia ArribaRESUMEN
BACKGROUND: Little is known about the onset and prevalence of periodontal disease in patients with the rare Kindler syndrome, a genodermatological disorder. This study investigated the level of clinical periodontal attachment in relation to age and presence of putative periodontopathogenic bacteria in individuals with Kindler syndrome. METHODS: Eighteen individuals diagnosed with Kindler syndrome and 13 control subjects, aged 4 to 37 years, from rural Panama received a limited clinical periodontal examination. Subgingival samples were collected for identification of putative periodontal pathogens by polymerase chain reaction. RESULTS: Mild to severe gingivitis was a common finding in all adults of the study population. Seventy-two percent (13/18) of the Kindler patients and 46% (6/13) of the control subjects showed mild to severe periodontal disease (P = 0.001, chi-square test). The onset of periodontitis was earlier and the progression occurred at a faster rate in the Kindler group. There was a strong correlation (r = 0.83) between the level of attachment loss and age in the Kindler group and a weaker correlation (r = 0.66) in the control group. The appearance of gingival tissues suggested atypical periodontitis with spontaneous bleeding and fragile, often desquamative, gingiva. In periodontitis patients, Porphyromonas gingivallis and Diallster pneumosintes tended to occur more frequently in control individuals compared to those with Kindler syndrome. CONCLUSIONS: In the Kindler group, periodontitis had an onset in early teenage years and progressed more rapidly compared to non-Kindler individuals of the same geographic and ethnic group. Clinical and microbiological findings suggest atypical periodontitis in Kindler patients. We propose to include Kindler syndrome in the category of medical disorders predisposing to destructive periodontal disease.
Asunto(s)
Periodontitis/complicaciones , Enfermedades Cutáneas Vesiculoampollosas/complicaciones , Adolescente , Adulto , Factores de Edad , Edad de Inicio , Anciano , Distribución de Chi-Cuadrado , Niño , Preescolar , Progresión de la Enfermedad , Susceptibilidad a Enfermedades , Epidermólisis Ampollosa/complicaciones , Epidermólisis Ampollosa/microbiología , Femenino , Hemorragia Gingival/complicaciones , Gingivitis/complicaciones , Bacilos Gramnegativos Anaerobios Rectos, Curvos y Espirales/aislamiento & purificación , Humanos , Masculino , Persona de Mediana Edad , Panamá , Pérdida de la Inserción Periodontal/complicaciones , Periodontitis/microbiología , Porphyromonas gingivalis/aislamiento & purificación , Síndrome Rothmund-Thomson/complicaciones , Síndrome Rothmund-Thomson/microbiología , Salud Rural , Enfermedades Cutáneas Vesiculoampollosas/microbiología , SíndromeRESUMEN
BACKGROUND: Measuring low back injury risk factors in field research presents challenges not encountered in laboratory environments. METHODS: We compared the practical application of five measurement methods (observations, interviews, electromyography (EMG), inclinometry, and vibration monitoring) for 223 worker days in 50 heavy-industry worksites in western Canada. Data collection successes, challenges, costs, and data detail were documented for each method. RESULTS: Measurement success rates varied from 42.2% (seatpan accelerometer) to 99.6% (post-shift interview) of worker days assessed. Missed days for direct monitoring equipment were primarily due to explosive environments, workplace conditions likely to damage the equipment, and malfunctions. Costs per successful measurement day were lowest for interviews (approximately 23 dollars), about 10-fold higher for observations and inclinometry, and more than 20-fold higher for EMG and vibration monitoring. CONCLUSIONS: Costs and successful field performance need to be weighed against the added data detail gained from monitoring equipment when making choices about exposure assessment techniques for epidemiological studies.