A high-content chemical screen identifies ellipticine as a modulator of p53 nuclear localization

p53 regulates apoptosis and the cell cycle through actions in the nucleus and cytoplasm. Altering the subcellular localization of p53 can alter its biological function. Therefore, small molecules that change the localization of p53 would be useful chemical probes to understand the influence of subcellular localization on the function of p53. To identify such molecules, a high-content screen for compounds that increased the localization of p53 to the nucleus or cytoplasm was developed, automated, and conducted. With this image-based assay, we identified ellipticine that increased the nuclear localization of GFP-mutant p53 protein but not GFP alone in Saos-2 osteosarcoma cells. In addition, ellipticine increased the nuclear localization of endogenous p53 in HCT116 colon cancer cells with a resultant increase in the transactivation of the p21 promoter. Increased nuclear p53 after ellipticine treatment was not associated with an increase in DNA double stranded breaks, indicating that ellipticine shifts p53 to the nucleus through a mechanism independent of DNA damage. Thus, a chemical biology approach has identified a molecule that shifts the localization of p53 and enhances its nuclear activity. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. G. Wei Xu and Imtiaz A. Mawji have contributed equally to this work.     

Identification and structural characterization of heme binding in a novel dye-decolorizing peroxidase, TyrA

TyrA is a member of the dye-decolorizing peroxidase (DyP) family, a new family of heme-dependent peroxidase recently identified in fungi and bacteria. Here, we report the crystal structure of TyrA in complex with iron protoporphyrin (IX) at 2.3 A. TyrA is a dimer, with each monomer exhibiting a two-domain, alpha/beta ferredoxin-like fold. Both domains contribute to the heme-binding site. Co-crystallization in the presence of an excess of iron protoporphyrin (IX) chloride allowed for the unambiguous location of the active site and the specific residues involved in heme binding. The structure reveals a Fe-His-Asp triad essential for heme positioning, as well as a novel conformation of one of the heme propionate moieties compared to plant peroxidases. Structural comparison to the canonical DyP family member, DyP from Thanatephorus cucumeris (Dec 1), demonstrates conservation of this novel heme conformation, as well as residues important for heme binding. Structural comparisons with representative members from all classes of the plant, bacterial, and fungal peroxidase superfamily demonstrate that TyrA, and by extension the DyP family, adopts a fold different from all other structurally characterized heme peroxidases. We propose that a new superfamily be added to the peroxidase classification scheme to encompass the DyP family of heme peroxidases.     

Convergent evolution of a labile nutritional symbiosis in ants

Ants are among the most successful organisms on Earth. It has been suggested that forming symbioses with nutrient-supplementing microbes may have contributed to their success, by allowing ants to invade otherwise inaccessible niches. However, it is unclear whether ants have evolved symbioses repeatedly to overcome the same nutrient limitations. Here, we address this question by comparing the independently evolved symbioses in Camponotus, Plagiolepis, Formica and Cardiocondyla ants. Our analysis reveals the only metabolic function consistently retained in all of the symbiont genomes is the capacity to synthesise tyrosine. We also show that in certain multi-queen lineages that have co-diversified with their symbiont for millions of years, only a fraction of queens carry the symbiont, suggesting ants differ in their colony-level reliance on symbiont-derived resources. Our results imply that symbioses can arise to solve common problems, but hosts may differ in their dependence on symbionts, highlighting the evolutionary forces influencing the persistence of long-term endosymbiotic mutualisms.Subject terms: Molecular evolution, Microbial ecology, Comparative genomics     

A transient apical extracellular matrix relays cytoskeletal patterns to shape permanent acellular ridges on the surface of adult C. elegans

Epithelial cells secrete apical extracellular matrices to form protruding structures such as denticles, ridges, scales, or teeth. The mechanisms that shape these structures remain poorly understood. Here, we show how the actin cytoskeleton and a provisional matrix work together to sculpt acellular longitudinal alae ridges in the cuticle of adult C. elegans. Transient assembly of longitudinal actomyosin filaments in the underlying lateral epidermis accompanies deposition of the provisional matrix at the earliest stages of alae formation. Actin is required to pattern the provisional matrix into longitudinal bands that are initially offset from the pattern of longitudinal actin filaments. These bands appear ultrastructurally as alternating regions of adhesion and separation within laminated provisional matrix layers. The provisional matrix is required to establish these demarcated zones of adhesion and separation, which ultimately give rise to alae ridges and their intervening valleys, respectively. Provisional matrix proteins shape the alae ridges and valleys but are not present within the final structure. We propose a morphogenetic mechanism wherein cortical actin patterns are relayed to the laminated provisional matrix to set up distinct zones of matrix layer separation and accretion that shape a permanent and acellular matrix structure.     

Effects of group dynamics and diet on the ranging patterns of a western gorilla group (Gorilla gorilla gorilla) at Bai Hokou, Central African Republic

This study describes how group dynamics and diet have influenced the ranging patterns of a western gorilla group at Bai Hokou, Central African Republic. The results are compared with those from an earlier study [Cipolletta, International Journal of Primatology, 2003], when the same group was larger and undergoing the process of habituation to humans. Data were obtained from maps of the gorillas' travel routes, direct observations, and analysis of fecal samples. Through the years, the group has experienced a decrease in size, from eight to three individuals, with periods of membership fluctuation. The male's search for new mates resulted in a larger home range than was recorded when the group consisted of more individuals. Moreover, despite an average group size of three throughout this study, the monthly range and mean daily path length (DPL) were also larger when the group was acquiring/losing members in new areas, than when no new members joined or left the group. Fruit was consumed year-round, although more heavily so during wet months. The influence of fruit consumption on the ranging patterns was concealed initially by the effect of habituation [Cipolletta, International Journal of Primatology, 2003], and later (at least partially) by the male's search for new mates. In the last 14 months of the study, when the group numbered only three individuals and was ranging in a restricted area, the average DPL, but not the monthly range, increased when the gorillas were consuming more fruit.     

KIR2DL2 enhances protective and detrimental HLA class I-mediated immunity in chronic viral infection

Killer cell immunoglobulin-like receptors (KIRs) influence both innate and adaptive immunity. But while the role of KIRs in NK-mediated innate immunity is well-documented, the impact of KIRs on the T cell response in human disease is not known. Here we test the hypothesis that an individual's KIR genotype affects the efficiency of their HLA class I-mediated antiviral immune response and the outcome of viral infection. We show that, in two unrelated viral infections, hepatitis C virus and human T lymphotropic virus type 1, possession of the KIR2DL2 gene enhanced both protective and detrimental HLA class I-restricted anti-viral immunity. These results reveal a novel role for inhibitory KIRs. We conclude that inhibitory KIRs, in synergy with T cells, are a major determinant of the outcome of persistent viral infection.     

The potential utility of carotenoid‐based coloration as a biomonitor of environmental change

In the past 30 years, carotenoid‐based animal signals have been an intense focus of research because they can potentially broadcast an honest reflection of individual reproductive potential. Our understanding of the underpinning physiological functions of carotenoid compounds is still emerging, however. Here, we argue that wildlife researchers and managers interested in assessing the impact of environmental quality on animal populations should be taking advantage of the signalling function of carotenoid‐based morphological traits. Using birds as a model taxonomic group, we build our argument by first reviewing the strong evidence that the expression of avian carotenoid displays provides an integrated measure of a multitude of diet‐ and health‐related parameters. We then present evidence that human‐induced rapid environmental change (HIREC) impacts the expression of carotenoid signals across different critical periods of a bird’s lifetime. Finally, we argue that variation in signal expression across individuals, populations and species could be quantified relatively easily at a global scale by incorporating such measurements into widespread bird ringing activities. Monitoring the expression of carotenoid‐based coloration could help to identify how the environmental factors linked to HIREC can affect avian populations and allow for potentially detrimental effects on biodiversity to be detected prior to demographic change.