Exploring the Nicotinic Acetylcholine Receptor-associated Proteome with iTRAQ and Transgenic Mice

Neuronal nicotinic acetylcholine receptors (nAChRs) containing α4 and β2 subunits are the principal receptors in the mammalian central nervous system that bind nicotine with high affinity. These nAChRs are involved in nicotine dependence, mood disorders, neurodegeneration and neuroprotection. However, our understanding of the interactions between α4β2-containing (α4β2¹) nAChRs and other proteins remains limited. In this study, we identified proteins that interact with α4β2¹ nAChRs in a genedose dependent pattern by immunopurifying β2¹ nAChRs from mice that differ in α4 and β2 subunit expression and performing proteomic analysis using isobaric tags for relative and absolute quantitation (iTRAQ). Reduced expression of either the α4 or the β2 subunit results in a correlated decline in the expression of a number of putative interacting proteins. We identified 208 proteins co-immunoprecipitated with these nAChRs. Furthermore, stratified linear regression analysis indicated that levels of 17 proteins was correlated significantly with expression of α4β2 nAChRs, including proteins involved in cytoskeletal rearrangement and calcium signaling. These findings represent the first application of quantitative proteomics to produce a β2¹ nAChR interactome and describe a novel technique used to discover potential targets for pharmacological manipulation of α4β2 nAChRs and their downstream signaling mechanisms.     

Correlates of Protective Cellular Immunity Revealed by Analysis of Population-Level Immune Escape Pathways in HIV-1

HLA class I-associated polymorphisms identified at the population level mark viral sites under immune pressure by individual HLA alleles. As such, analysis of their distribution, frequency, location, statistical strength, sequence conservation, and other properties offers a unique perspective from which to identify correlates of protective cellular immunity. We analyzed HLA-associated HIV-1 subtype B polymorphisms in 1,888 treatment-naïve, chronically infected individuals using phylogenetically informed methods and identified characteristics of HLA-associated immune pressures that differentiate protective and nonprotective alleles. Over 2,100 HLA-associated HIV-1 polymorphisms were identified, approximately one-third of which occurred inside or within 3 residues of an optimally defined cytotoxic T-lymphocyte (CTL) epitope. Differential CTL escape patterns between closely related HLA alleles were common and increased with greater evolutionary distance between allele group members. Among 9-mer epitopes, mutations at HLA-specific anchor residues represented the most frequently detected escape type: these occurred nearly 2-fold more frequently than expected by chance and were computationally predicted to reduce peptide-HLA binding nearly 10-fold on average. Characteristics associated with protective HLA alleles (defined using hazard ratios for progression to AIDS from natural history cohorts) included the potential to mount broad immune selection pressures across all HIV-1 proteins except Nef, the tendency to drive multisite and/or anchor residue escape mutations within known CTL epitopes, and the ability to strongly select mutations in conserved regions within HIV''s structural and functional proteins. Thus, the factors defining protective cellular immune responses may be more complex than simply targeting conserved viral regions. The results provide new information to guide vaccine design and immunogenicity studies.     

A novel founder mutation in the RNASEL gene, 471delAAAG,is associated with prostate cancer in Ashkenazi Jews

HPC1/RNASEL was recently identified as a candidate gene for hereditary prostate cancer. We identified a novel founder frameshift mutation in RNASEL, 471delAAAG, in Ashkenazi Jews. The mutation frequency in the Ashkenazi population, estimated on the basis of the frequency in 150 healthy young women, was 4% (95% confidence interval [CI] 1.9%-8.4%). Among Ashkenazi Jews, the mutation frequency was higher in patients with prostate cancer (PRCA) than in elderly male control individuals (6.9% vs. 2.4%; odds ratio = 3.0; 95% CI 0.6-15.3; P=.17). 471delAAAG was not detected in the 134 non-Ashkenazi patients with PRCA and control individuals tested. The median age at PRCA diagnosis did not differ significantly between the Ashkenazi carriers and noncarriers included in our study. However, carriers received diagnoses at a significantly earlier age, compared with patients with PRCA who were registered in the Israeli National Cancer Registry (65 vs. 74.4 years, respectively; P<.001). When we examined two brothers with PRCA, we found a heterozygous 471delAAAG mutation in one and a homozygous mutation in the other. Loss of heterozygosity was demonstrated in the tumor of the heterozygous sib. Taken together, these data suggest that the 471delAAAG null mutation is associated with PRCA in Ashkenazi men. However, additional studies are required to determine whether this mutation confers increased risk for PRCA in this population.     

Reproductive biology of the sand dollar,Peronella lesueuri (L. Agassiz, 1841) (Echinoidea: Clypeasteroida: Laganidae), in south-western Australia

The reproductive biology of the sand dollar Peronella lesueuri was studied between 2009 and 2011 in Cockburn Sound, a large coastal embayment in south-western Australia. Individuals of P. lesueuri did not display sexual size-dimorphism, and the population was found to have a sex ratio of 1:1. Maturity occurred over the range of 80–115 mm with all sand dollars larger than 115 mm having distinct gonads. Monthly histological analysis of gonads and changes in oocyte proportions over time indicated that P. lesueuri has an annual reproductive cycle; gametogenesis occurs in spring and spawning in summer. Differences in the rate of gametogenesis between 2009/2010 and 2010/2011 may have been influenced by higher temperatures experienced in 2010/2011. P. lesueuri have large ova (mean?=?210 μm), which suggests the species has lecithotrophic larval development.     

Den attendance patterns in swift foxes during pup rearing: varying degrees of parental investment within the breeding pair

Parental investment varies in mammalian species, with male care of young being more common in social and monogamous species. Monogamy is commonly observed in canid species, with both males and females, and often “helper” individuals, providing some degree of care for the young. Social units of the swift fox (Vulpes velox), a small North American canid species, usually consist of a male–female pair and occasionally helpers. The role of parental investment and behavior in swift fox society is currently poorly understood. We observed swift fox dens during the pup-rearing season in each of 2 years to evaluate attendance and frequency of visits to natal dens by adult males and females. Female foxes remained at dens longer and visited them more frequently than did male foxes. Female attendance and visitation decreased throughout the pup-rearing season as pups became older and more independent. Environmental factors, including climate and its effect on prey, appeared to contribute to differences in fox behavior between the 2 years. We observed only one fox outside of the breeding pair attending a den in each of the 2 years, both of which were males. We concluded that each of these two foxes were living within the social unit of the male–female pair as a trio, but not serving as a helper and contributing to the care of the pups. Our results increased knowledge of the ecology and behavior of the swift fox, a species of conservation concern in the Great Plains of North America.     

Unemployment and labor migration in rural Galicia (Spain)

This article focuses on the difficulty of entering into—and literally getting back and forth to—the waged labor market for adolescent and young adult workers from rural parts of Galicia, a region that has experienced a prolonged history of persistently high levels of unemployment. Unemployment is one of the most serious issues facing Spain, other countries of the EU, and states all over the world. In Spain, for the population of workers under 30, this problem is at staggeringly high levels that have reached 50 % plus in the last few years of the current crisis but has been longstanding. So, too, has labor migration to and from rural Galician households. This article takes a broad approach to the analysis of labor mobility, including daily commuting as well as longer distance and longer duration labor migration. It considers the role of state and EU policies over time in instituting, supporting, or discouraging different geographical trajectories and forms of movement for young adult “wage seekers” from rural Galician households.     

Structural studies of p53 inactivation by DNA-contact mutations and its rescue by suppressor mutations via alternative protein–DNA interactions

A p53 hot-spot mutation found frequently in human cancer is the replacement of R273 by histidine or cysteine residues resulting in p53 loss of function as a tumor suppressor. These mutants can be reactivated by the incorporation of second-site suppressor mutations. Here, we present high-resolution crystal structures of the p53 core domains of the cancer-related proteins, the rescued proteins and their complexes with DNA. The structures show that inactivation of p53 results from the incapacity of the mutated residues to form stabilizing interactions with the DNA backbone, and that reactivation is achieved through alternative interactions formed by the suppressor mutations. Detailed structural and computational analysis demonstrates that the rescued p53 complexes are not fully restored in terms of DNA structure and its interface with p53. Contrary to our previously studied wild-type (wt) p53-DNA complexes showing non-canonical Hoogsteen A/T base pairs of the DNA helix that lead to local minor-groove narrowing and enhanced electrostatic interactions with p53, the current structures display Watson–Crick base pairs associated with direct or water-mediated hydrogen bonds with p53 at the minor groove. These findings highlight the pivotal role played by R273 residues in supporting the unique geometry of the DNA target and its sequence-specific complex with p53.     

“Frame That Rug!”: Narratives of Zapotec Textiles as Art and Ethnic Commodity in the Global Marketplace

This article explores representations of Mexican Zapotec textiles as texts and as sites of contested values and meanings. Discussion of this indigenous cottage industry begins in the Oaxacan village of Teotitlán del Valle, the site of production, and follows the textile as it moves across social and cultural borders as a commodity in postindustrial consumer culture. The anthropologist's mediating role in brokering Zapotec textiles is considered within the social context of exchange relations between buyer and seller. Ethnographic stories about the production and aesthetics of Zapotec textiles are contrasted with consumers’ self-fashioning narratives of embodied experiences associated with their hand-woven possessions. The article concludes with reflections on the narrative threads of text that comprise a Zapotec textile's intrinsic value as both art and craft.