Biochemical and biophysical characterization of the RVB-1/RVB-2 protein complex, the RuvBL/RVB homologues in Neurospora crassa.

The RVB proteins, composed of the conservative paralogs, RVB1 and RVB2, belong to the AAA+ (ATPases Associated with various cellular Activities) protein superfamily and are present in archaea and eukaryotes. The most distinct structural features are their ability to interact with each other forming the RVB1/2 complex and their participation in several macromolecular protein complexes leading them to be involved in many biological processes. We report here the biochemical and biophysical characterization of the Neurospora crassa RVB-1/RVB-2 complex. Chromatographic analyses revealed that the complex (APO) predominantly exists as a dimer in solution although hexamers were also observed. Nucleotides influence the oligomerization state, while ATP favors hexamers formation, ADP favors the formation of multimeric states, likely dodecamers, and the Molecular Dynamics (MD) simulations revealed the contribution of certain amino acid residues in the nucleotide stabilization. The complex binds to dsDNA fragments and exhibits ATPase activity, which is strongly enhanced in the presence of DNA. In addition, both GFP-fused proteins are predominantly nuclear, and their nuclear localization signals (NLS) interact with importin-α (NcIMPα). Our findings show that some properties are specific of the fungus proteins despite of their high identity to orthologous proteins. They are essential proteins in N. crassa, and the phenotypic defects exhibited by the heterokaryotic strains, mainly related to growth and development, indicate N. crassa as a promising organism to investigate additional biological and structural aspects of these proteins.

Photoelastic stress analysis of different types of anterior teeth splints.

BACKGROUND/AIMS: Traumatic dental injuries are highly prevalent and are considered an important health problem. The aim of this study was to evaluate the in vitro stress distribution around simulated traumatized teeth subjected to different types of splints by photoelastic analysis. MATERIALS AND METHODS: For each group, five models of maxillary and mandibular arches were made in photoelastic resin using prefabricated teeth models, which were fixed and adjusted in a semi-adjustable articulator. All splints were bonded with composite resin on the labial surfaces of the maxillary central incisors, the lateral incisors and the canine teeth, simulating trauma to the 11. The groups were divided according to the type of splint: control-without splint; rigid-brackets and 0.021 × 0.025 inch stainless steel wire; semi-rigid-brackets and 0.016 × 0.016 inch cobalt-chromium wire; fiber-fiberglass ribbon; and flexible-nylon thread. The groups were submitted to an occlusal force in a special device attached to a universal test machine in balanced, protrusive, and lateral occlusions. Then, the incisal edge of the 11 was ground to simulate infra-occlusion, and the tests were performed again. Five points were analyzed around tooth 11 with a polariscope, and the data were submitted to ANOVA and Tukey tests at a 5% level of significance. RESULTS: When centric occlusion was analyzed by comparing normal occlusion to infra-occlusion, all groups showed high stress values in infra-occlusion with statistical differences-except for the control group. When lateral occlusion was analyzed, the nylon splint showed lower statistical differences compared with the control, fiberglass, and rigid splint groups, which did not differ between each other. The semi-rigid splint did not differ from any group. CONCLUSIONS: Fiberglass and semi-rigid splints showed better stress distribution around the traumatized tooth under occlusal loads. The nylon splint showed characteristics of non-stress distribution.