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Jose Maria Carazo

Publications and source records attributed to Jose Maria Carazo.

3 recordsLinked to original sources

On the nature of 2D crystal unbending.

Crystal unbending, the process that aims to recover a perfect crystal from experimental data, is one of the more important steps in electron crystallography image processing. The unbending process involves three steps: estimation of the unit cell displacements from their ideal positions, extension of the deformation field to the whole image and transformation of the image in order to recover an ideal crystal. In this work, we present a systematic analysis of the second step oriented to address two issues. First, whether the unit cells remain undistorted and only the distance between them should be changed (rigid case) or should be modified with the same deformation suffered by the whole crystal (elastic case). Second, the performance of different extension algorithms (interpolation versus approximation) is explored. Our experiments show that there is no difference between elastic and rigid cases or among the extension algorithms. This implies that the deformation fields are constant over large areas. Furthermore, our results indicate that the main source of error is the transformation of the crystal image.

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Large T antigen on the simian virus 40 origin of replication: a 3D snapshot prior to DNA replication.

Large T antigen is the replicative helicase of simian virus 40. Its specific binding to the origin of replication and oligomerization into a double hexamer distorts and unwinds dsDNA. In viral replication, T antigen acts as a functional homolog of the eukaryotic minichromosome maintenance factor MCM. T antigen is also an oncoprotein involved in transformation through interaction with p53 and pRb. We obtained the three-dimensional structure of the full-length T antigen double hexamer assembled at its origin of replication by cryoelectron microscopy and single-particle reconstruction techniques. The double hexamer shows different degrees of bending along the DNA axis. The two hexamers are differentiated entities rotated relative to each other. Isolated strands of density, putatively assigned to ssDNA, protrude from the hexamer-hexamer junction mainly at two opposite sites. The structure of the T antigen at the origin of replication can be understood as a snapshot of the dynamic events leading to DNA unwinding. Based on these results a model for the initiation of simian virus 40 DNA replication is proposed.

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