The secreted proteins of Achlya hypogyna and Thraustotheca clavata identify the ancestral oomycete secretome and reveal gene acquisitions by horizontal gene transfer

This is the final version. Available on open access from Oxford University Press via the DOI in this recordSaprotrophic and parasitic microorganisms secrete proteins into the environment to breakdown macromolecules and obtain nutrients. The molecules secreted are collectively termed the "secretome" and the composition and function of this set of proteins varies depending on the ecology, life cycle, and environment of an organism. Beyond the function of nutrient acquisition, parasitic lineages must also secrete molecules to manipulate their host. Here, we use a combination of de novo genome and transcriptome sequencing and bioinformatic identification of signal peptides to identify the putative secreted proteome of two oomycetes, the facultative parasite Achlya hypogyna and free-living Thraustotheca clavata. By comparing the secretomes of these saprolegnialean oomycetes with that of eight other oomycetes, we were able to characterize the evolution of this protein set across the oomycete clade. These species span the last common ancestor of the two major oomycete families allowing us to identify the ancestral secretome. This putative ancestral secretome consists of at least 84 gene families. Only 11 of these gene families are conserved across all 10 secretomes analyzed and the two major branches in the oomycete radiation. Notably, we have identified expressed elicitin-like effector genes in the saprotrophic decomposer, T. clavata. Phylogenetic analyses show six novel horizontal gene transfers to the oomycete secretome from bacterial and fungal donor lineages, four of which are specific to the Saprolegnialeans. Comparisons between free-living and pathogenic taxa highlight the functional changes of oomycete secretomes associated with shifts in lifestyle.University of Rhode Island College of the Environment and Life SciencesU.S. Department of AgricultureNational Institutes of Health (NIH)National Science FoundationDivision of Biology and Medicine, Brown Universit

Spherulitic crystal growth drives mineral deposition patterns in collagen-based materials

The formation of the hard tissues that provide support and mobility to organisms is achieved through the interplay of inorganic crystals and an organic framework composed of collagen and a small percentage of non-collagenous proteins. Despite their clinical relevance, the mechanisms governing mineralization of the extracellular matrix are still poorly understood. By using 3D electron tomography and high-resolution electron microscopy imaging and spectroscopy, it has been demonstrated that mineralization proceeds through a spherulitic-like crystal growth process. First, aggregates of disordered crystals form in the interfibrillar spaces, which lead to the mineralization of adjacent fibrils. Mineral propagates steadily through the inter- and intrafibrillar spaces of the collagen structure forming layered spherulites that grow to confluence. The structure of the collagen fibrils serves as a protein scaffold to guide the formation of a myriad of platelet-shaped crystallites that make up each of these spherulites. At their periphery, nanosized unmineralized areas remain, leading to the formation of the characteristic lacy pattern observed in the transversal cross-section of mature calcified tissues. This study provides fundamental insights into the bone formation process and represents a potential strategy for complex materials design

Molecular characterisation of viruses from Kiwifruit

In 2003 Apple stem grooving virus was discovered in Actinidia accessions from China, being held in quarantine in Auckland. Subsequent examination of kiwifruit germplasm from the same source has detected several additional viruses, including a ~300 nm rigid rod related to Ribgrass mosaic virus (Tobamovirus), a 700-750 nm flexuous virus related to Citrus leaf blotch virus (Flexiviridae) and a novel vitivirus. Currently these viruses have not been reported from commercial kiwifruit crops in New Zealand or elsewhere. The biological properties of the viruses from kiwifruit and their phylogenetic relationships with similar viruses from other plants will be described, and the possible implications for the international movement of Actinidia germplasm are discussed

The lipoatrophic caveolin-1 deficient mouse model reveals autophagy in mature adipocytes

Adipose tissue lipoatrophy caused by caveolin gene deletion in mice is not linked to defective adipocyte differentiation. We show that adipose tissue development cannot be rescued by endothelial specific caveolin-1 re-expression, indicating primordial role of caveolin in mature adipocytes. Partial or total caveolin deficiency in adipocytes induced broad protein expression defects, including but not limited to previously described downregulation of insulin receptor. Global alterations in protein turnover, and accelerated degradation of long-lived proteins were found in caveolin-deficient adipocytes. Lipidation of endogenous LC3 autophagy marker and distribution of GFP-LC3 into aggregates demonstrated activated autophagy in the absence of caveolin-1 in adipocytes. Furthermore, electron microscopy revealed autophagic vacuoles in caveolin-1 deficient but not control adipocytes. Surprisingly, significant levels of lipidated LC3-II were found around lipid droplets of normal adipocytes, maintained in nutrient-rich conditions or isolated from fed mice, which do not display autophagy. Altogether, these data indicate that caveolin deficiency induce autophagy in adipocytes, a feature that is not a physiological response to fasting in normal fat cells. This likely resulted from defective insulin and lipolytic responses that converge in chronic nutrient shortage in adipocytes lacking caveolin-1. This is the first report of a pathological situation with autophagy as an adaptative response to adipocyte failure

Nondestructive Structure Characterization by Laser-Ultrasonics

Peer reviewed: YesNRC publication: Ye

Quantitative backscattered electron imaging of bone using a thermionic or a field emission electron source

Quantitative backscattered electron imaging is an established method to map mineral content distributions in bone and to determine the bone mineralization density distribution (BMDD). The method we applied was initially validated for a scanning electron microscope (SEM) equipped with a tungsten hairpin cathode (thermionic electron emission) under strongly defined settings of SEM parameters. For several reasons, it would be interesting to migrate the technique to a SEM with a field emission electron source (FE-SEM), which, however, would require to work with different SEM parameter settings as have been validated for DSM 962. The FE-SEM has a much better spatial resolution based on an electron source size in the order of several 100 nanometers, corresponding to an about 105 to 106 times smaller source area compared to thermionic sources. In the present work, we compare BMDD between these two types of instruments in order to further validate the methodology. We show that a transition to higher pixel resolution (1.76, 0.88, and 0.57 μm) results in shifts of the BMDD peak and BMDD width to higher values. Further the inter-device reproducibility of the mean calcium content shows a difference of up to 1 wt% Ca, while the technical variance of each device can be reduced to ±0.17 wt% Ca. Bearing in mind that shifts in calcium levels due to diseases, e.g., high turnover osteoporosis, are often in the range of 1 wt% Ca, both the bone samples of the patients as well as the control samples have to be measured on the same SEM device. Therefore, we also constructed new reference BMDD curves for adults to be used for FE-SEM data comparison