RESUMEN
Translocation of nuclear-encoded preproteins across the inner envelope of chloroplasts is catalyzed by the Tic translocon, consisting of Tic110, Tic40, Tic62, Tic55, Tic32, Tic20, and Tic22. Tic62 was proposed to act as a redox sensor of the complex because of its redox-dependent shuttling between envelope and stroma and its specific interaction with the photosynthetic protein ferredoxin-NADP(H) oxidoreductase (FNR). However, the nature of this close relationship so far remained enigmatic. A putative additional localization of Tic62 at the thylakoids mandated further studies examining how this feature might be involved in the respective redox sensing pathway and the interaction with its partner protein. Therefore, both the association with FNR and the physiological role of the third, thylakoid-bound pool of Tic62 were investigated in detail. Coexpression analysis indicates that Tic62 has similar expression patterns as genes involved in photosynthetic functions and protein turnover. At the thylakoids, Tic62 and FNR form high molecular weight complexes that are not involved in photosynthetic electron transfer but are dynamically regulated by light signals and the stromal pH. Structural analyses reveal that Tic62 binds to FNR in a novel binding mode for flavoproteins, with a major contribution from hydrophobic interactions. Moreover, in absence of Tic62, membrane binding and stability of FNR are drastically reduced. We conclude that Tic62 represents a major FNR interaction partner not only at the envelope and in the stroma, but also at the thylakoids of Arabidopsis thaliana and perhaps all flowering plants. Association with Tic62 stabilizes FNR and is involved in its dynamic and light-dependent membrane tethering.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Cloroplastos/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Oxidorreductasas/metabolismo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , ADN Bacteriano/genética , Regulación de la Expresión Génica de las Plantas , Técnicas de Inactivación de Genes , Luz , Proteínas de Transporte de Membrana/genética , Modelos Moleculares , Mutagénesis Insercional , Mutación , Análisis de Secuencia por Matrices de Oligonucleótidos , Oxidación-Reducción , Oxidorreductasas/genética , Fotosíntesis , Unión Proteica , Mapeo de Interacción de Proteínas , ARN de Planta/genéticaRESUMEN
In order to further evaluate the process of microwear formation on human dental enamel, microwear was experimentally produced by a chewing simulation with an Academic Center for Dentistry Amsterdam (ACTA) device. For this simulation, several cereal species were processed according to historical milling techniques, the experimental results of which were compared with those obtained from cereals processed after modern techniques, and also with natural microwear on early medieval human molars. Comparison of simulated microwear pits with natural microwear pits showed that the simulation led to traces which matched those found on the historical teeth in terms of both size and shape. Experimentally produced microwear pits were especially characteristic for the cereal species used in the simulations, and both pit morphology and enamel loss were a function of cereal phytolith content. Despite the high variability of phytolith size and shape, certain types are characteristic for certain cereals, which in turn are capable of producing cereal-specific microwear. This experimental approach is likely to further define ancient human dietary behavior, including food processing.