Studying Coxiella burnetii Type IV Substrates in the Yeast Saccharomyces cerevisiae: Focus on Subcellular Localization and Protein Aggregation.

Coxiella burnetii is a Gram-negative obligate parasitic bacterium that causes the disease Q-fever in humans. To establish its intracellular niche, it utilizes the Icm/Dot type IVB secretion system (T4BSS) to inject protein effectors into the host cell cytoplasm. The host targets of most cognate and candidate T4BSS-translocated effectors remain obscure. We used the yeast Saccharomyces cerevisiae as a model to express and study six C. burnetii effectors, namely AnkA, AnkB, AnkF, CBU0077, CaeA and CaeB, in search for clues about their role in C. burnetii virulence. When ectopically expressed in HeLa cells, these effectors displayed distinct subcellular localizations. Accordingly, GFP fusions of these proteins produced in yeast also decorated distinct compartments, and most of them altered cell growth. CaeA was ubiquitinated both in yeast and mammalian cells and, in S. cerevisiae, accumulated at juxtanuclear quality-control compartments (JUNQs) and insoluble protein deposits (IPODs), characteristic of aggregative or misfolded proteins. AnkA, which was not ubiquitinated, accumulated exclusively at the IPOD. CaeA, but not AnkA or the other effectors, caused oxidative damage in yeast. We discuss that CaeA and AnkA behavior in yeast may rather reflect misfolding than recognition of conserved targets in the heterologous system. In contrast, CBU0077 accumulated at vacuolar membranes and abnormal ER extensions, suggesting that it interferes with vesicular traffic, whereas AnkB associated with the yeast nucleolus. Both effectors shared common localization features in HeLa and yeast cells. Our results support the idea that C. burnetii T4BSS effectors manipulate multiple host cell targets, which can be conserved in higher and lower eukaryotic cells. However, the behavior of CaeA and AnkA prompt us to conclude that heterologous protein aggregation and proteostatic stress can be a limitation to be considered when using the yeast model to assess the function of bacterial effectors.

Architecture of a Bench-Type Carbonate Lake Margin and Its Relation to Fluvially Dominated Deltas, Las Minas Basin, Upper Miocene, Spain

The Upper Miocene stratigraphic succession of the Las Minas Basin, located at the external zone of the Betic Chain in SE Spain, preserves several examples of lake carbonate bench deposits. Excellent exposures of the carbonate benches allow detailed observation of the architecture of these sediments and provide new insights for the ‘‘steep-gradient bench margin–low energy’’ model proposed by Platt and Wright (1991). The lake carbonate benches developed in close association with fluvially dominated shallow deltas that exhibit typical Gilbert-type profiles. The delta sequences comprise bottomset prodelta marl facies, distal to proximal foreset facies, deposited mainly in a delta-front environment, and topset facies, the latter reflecting both subaqueous delta-front and subaerial delta-plain environments. The development of the carbonate benches was constrained by the convexupward morphology of the deltaic deposits, which led to the available accommodation space for the growth of the steep-gradient platforms. The benches display a progradational pattern characterized by sigmoid-oblique internal geometries and offlap upper boundary relationships, which suggests that the carbonate benches developed under slow though continuous lake-level rise. Both the dimensions of the benches and the dominant carbonate components (i.e., encrusted charophyte stems and calcified cyanobaterial remains), allow comparisons with the progradational marl benches recognized in modern temperate hardwater lakes. Accordingly, the case study presented here provides a good ancient sedimentary analog for low-energy lake carbonate benches. Moreover, the evolutionary trend inferred from the fossil example offers new insights into the depositional conditions of this type of sediment and allows recognition of the transitional pattern from bench to ramp carbonate lake margins.

Cell Pattern in Adult Human Corneal Endothelium

A review of the current data on the cell density of normal adult human endothelial cells was carried out in order to establish some common parameters appearing in the different considered populations. From the analysis of cell growth patterns, it is inferred that the cell aging rate is similar for each of the different considered populations. Also, the morphology, the cell distribution and the tendency to hexagonallity are studied. The results are consistent with the hypothesis that this phenomenon is analogous with cell behavior in other structures such as dry foams and grains in polycrystalline materials. Therefore, its driving force may be controlled by the surface tension and the mobility of the boundaries.