Dynamic effects of cofactors and DNA on the oligomeric state of human mitochondrial DNA helicase

We examined the effects of cofactors and DNA on the stability, oligomeric state and conformation of the human mitochondrial DNA helicase. We demonstrate that low salt conditions result in protein aggregation that may cause dissociation of oligomeric structure. The low salt sensitivity of the mitochondrial DNA helicase is mitigated by the presence of magnesium, nucleotide, and increased temperature. Electron microscopic and glutaraldehyde cross-linking analyses provide the first evidence of a heptameric oligomer and its interconversion from a hexameric form. Limited proteolysis by trypsin shows that binding of nucleoside triphosphate produces a conformational change that is distinct from the conformation observed in the presence of nucleoside diphosphate. We find that single-stranded DNA binding occurs in the absence of cofactors and renders the mitochondrial DNA helicase more susceptible to proteolytic digestion. Our studies indicate that the human mitochondrial DNA helicase shares basic properties with the SF4 replicative helicases, but also identify common features with helicases outside the superfamily, including dynamic conformations similar to other AAA⁺ ATPases.

Using a brief questionnaire to assess the overall perception of basketball spectators

Purpose – The purpose of this paper is twofold. The first aim is to obtain a valid and reliable instrument for the holistic analysis of sporting events, and the second is to test a causal model in which future intentions depend on spectators’ perceptions of quality, satisfaction, and value of these events. Design/methodology/approach – A total of 493 spectators of a professional basketball team in the Spanish ACB league responded to a survey to measure the overall performance of the sporting event service. Exploratory factor analysis and further confirmatory factor analysis using structural equation models provides the methodology for testing the reliability and validity of the instrument. Findings – The scales have adequate reliability and validity indices. The path model explains 35.8 percent of the variance in future intentions, 54.0 percent in perceived value, and 49.5 percent in spectators’ satisfaction. Quality proves a better predictor of perceived value than satisfaction. Both perceived value and satisfaction have a similar weight in predicting spectators’ future intentions. The data indicate that quality has an effect on spectators’ future intentions, by altering their perceptions of value and satisfaction. Research limitations/implications – The research findings are somewhat limited, due to the sample consisting entirely of spectators of a single team in the Spanish ACB league. Practical implications – Managers can use these findings to develop loyalty strategies by creating service value and increasing spectators’ satisfaction through quality improvements. Originality/value – This study contributes to the literature on service quality by providing an overall measure to assess service in professional sporting events in a Latin-American context.