XDS

The paper describes the software package XDS for processing of single crystal diffraction data recorded by the rotation method

In-situ investigations of structural changes during cyclic loading by high resolution reciprocal space mapping

A major failure reason for structural materials is fatigue-related damage due to repeatedly changing mechanical loads. During cyclic loading dislocations self-organize into characteristic ordered structures, which play a decisive role for the materials lifetime. These heterogeneous dislocation structures can be identified using advanced electron microscopy and synchrotron techniques. A detailed characterization of the microstructure during cyclic loading by in-situ monitoring the internal structure within individual grains with high energy x-rays can help to understand and predict the materials behavior during cyclic deformation and to improve the material design. While monitoring macroscopic stress and strain during cyclic loading, reciprocal space maps of diffraction peaks from single grains are obtained with high resolution. High Resolution Reciprocal Space Mapping was applied successfully in-situ during cyclic deformation of macroscopic aluminium samples at the Advanced Photon Source to reveal the structural reorganization within single grains embedded in the bulk material during fatigue

Monitoring microstructural evolution in-situ during cyclic deformation by high resolution reciprocal space mapping

The recently developed synchrotron technique High Resolution Reciprocal Space Mapping (HRRSM) is used to characterize the deformation structures evolving during cyclic deformation of commercially pure, polycrystalline aluminium AA1050. Insight into the structural reorganization within single grains is gained by in-situ monitoring of the microstructural evolution during cyclic deformation. By HRRSM, a large number of individual subgrains can be resolved within individual grains in the bulk of polycrystalline specimens and their fate, their individual orientation and elastic stresses, tracked during different loading regimes as tension and compression. With this technique, the evolution of dislocation structures in selected grains was followed during an individual load cycle

Integration, scaling, space-group assignment and post-refinement

The working principles of important steps in processing rotation data are described as employed by the program XDS

The Inhibitor of Growth Protein 5 (ING5) Depends on INCA1 as a Co-Factor for Its Antiproliferative Effects

The proteins of the Inhibitor of Growth (ING) family are involved in multiple cellular functions such as cell cycle regulation, apoptosis, and chromatin remodeling. For ING5, its actual role in growth suppression and the necessary partners are not known. In a yeast-two-hybrid approach with human bone marrow derived cDNA, we identified ING5 as well as several other proteins as interaction partners of Inhibitor of cyclin A1 (INCA1) that we previously characterized as a novel interaction partner of cyclin A1/CDK2. ING5 expression in leukemic AML blasts was severely reduced compared to normal bone marrow. In line, ING5 inhibited bone marrow colony formation upon retroviral transduction. However, Inca1−/− bone marrow colony formation was not suppressed by ING5. In murine embryonic fibroblast (MEF) cells from Inca1+/+ and Inca1−/− mice, overexpression of ING5 suppressed cell proliferation only in the presence of INCA1, while ING5 had no effect in Inca1−/− MEFs. ING5 overexpression induced a delay in S-phase progression, which required INCA1. Finally, ING5 overexpression enhanced Fas-induced apoptosis in Inca1+/+ MEFs, while Inca1−/− MEFs were protected from Fas antibody-induced apoptosis. Taken together, these results indicate that ING5 is a growth suppressor with suppressed expression in AML whose functions depend on its interaction with INCA1

A protein functional leap: how a single mutation reverses the function of the transcription regulator TetR

Today's proteome is the result of innumerous gene duplication, mutagenesis, drift and selection processes. Whereas random mutagenesis introduces predominantly only gradual changes in protein function, a case can be made that an abrupt switch in function caused by single amino acid substitutions will not only considerably further evolution but might constitute a prerequisite for the appearance of novel functionalities for which no promiscuous protein intermediates can be envisaged. Recently, tetracycline repressor (TetR) variants were identified in which binding of tetracycline triggers the repressor to associate with and not to dissociate from the operator DNA as in wild-type TetR. We investigated the origin of this activity reversal by limited proteolysis, CD spectroscopy and X-ray crystallography. We show that the TetR mutant Leu17Gly switches its function via a disorder–order mechanism that differs completely from the allosteric mechanism of wild-type TetR. Our study emphasizes how single point mutations can engender unexpected leaps in protein function thus enabling the appearance of new functionalities in proteins without the need for promiscuous intermediates