The B55α Regulatory Subunit of Protein Phosphatase 2A Mediates Fibroblast Growth Factor-Induced p107 Dephosphorylation and Growth Arrest in Chondrocytes

Fibroblast growth factor (FGF)-induced growth arrest of chondrocytes is a unique cell type-specific response which contrasts with the proliferative response of most cell types and underlies several genetic skeletal disorders caused by activating FGF receptor (FGFR) mutations. We have shown that one of the earliest key events in FGF-induced growth arrest is dephosphorylation of the retinoblastoma protein (Rb) family member p107 by protein phosphatase 2A (PP2A), a ubiquitously expressed multisubunit phosphatase. In this report, we show that the PP2A-B55α holoenzyme (PP2A containing the B55α subunit) is responsible for this phenomenon. Only the B55α (55-kDa regulatory subunit, alpha isoform) regulatory subunit of PP2A was able to bind p107, and this interaction was induced by FGF in chondrocytes but not in other cell types. Small interfering RNA (siRNA)-mediated knockdown of B55α prevented p107 dephosphorylation and FGF-induced growth arrest of RCS (rat chondrosarcoma) chondrocytes. Importantly, the B55α subunit bound with higher affinity to dephosphorylated p107. Since the p107 region interacting with B55α is also the site of cyclin-dependent kinase (CDK) binding, B55α association may also prevent p107 phosphorylation by CDKs. FGF treatment induces dephosphorylation of the B55α subunit itself on several serine residues that drastically increases the affinity of B55α for the PP2A A/C dimer and p107. Together these observations suggest a novel mechanism of p107 dephosphorylation mediated by activation of PP2A through B55α dephosphorylation. This mechanism might be a general signal transduction pathway used by PP2A to initiate cell cycle arrest when required by external signals.     

Physico-chemical properties of R140G and K141Q mutants of human small heat shock protein HspB1 associated with hereditary peripheral neuropathies

Some physico-chemical properties of R140G and K141Q mutants of human small heat shock protein HspB1 associated with hereditary peripheral neuropathy were analyzed. Mutation K141Q did not affect intrinsic Trp fluorescence and interaction with hydrophobic probe bis-ANS, whereas mutation R140G decreased both intrinsic fluorescence and fluorescence of bis-ANS bound to HspB1. Both mutations decreased thermal stability of HspB1. Mutation R140G increased, whereas mutation K141Q decreased the rate of trypsinolysis of the central part (residues 5–188) of HspB1. Both the wild type HspB1 and its K141Q mutant formed large oligomers with apparent molecular weight ∼560 kDa. The R140G mutant formed two types of oligomers, i.e. large oligomers tending to aggregate and small oligomers with apparent molecular weight ∼70 kDa. The wild type HspB1 formed mixed homooligomers with R140G mutant with apparent molecular weight ∼610 kDa. The R140G mutant was unable to form high molecular weight heterooligomers with HspB6, whereas the K141Q mutant formed two types of heterooligomers with HspB6. In vitro measured chaperone-like activity of the wild type HspB1 was comparable with that of K141Q mutant and was much higher than that of R140G mutant. Mutations of homologous hot-spot Arg (R140G of HspB1 and R120G of αB-crystallin) induced similar changes in the properties of two small heat shock proteins, whereas mutations of two neighboring residues (R140 and K141) induced different changes in the properties of HspB1.     

Statistical torsion angle potential energy functions for protein structure modeling: A bicubic interpolation approach

A set of grid type knowledge‐based energy functions is introduced for ?–χ₁, ψ–χ₁, ?–ψ, and χ₁–χ₂ torsion angle combinations. Boltzmann distribution is assumed for the torsion angle populations from protein X‐ray structures, and the functions are named as statistical torsion angle potential energy functions. The grid points around periodic boundaries are duplicated to force periodicity, and the remedy relieves the derivative discontinuity problem. The devised functions rapidly improve the quality of model structures. The potential bias in the functions and the usefulness of additional secondary structure information are also investigated. The proposed guiding functions are expected to facilitate protein structure modeling, such as protein structure prediction, protein design, and structure refinement. Proteins 2013. Proteins 2013; 81:1156–1165. © 2013 Wiley Periodicals, Inc.     

Does Physical Contact with a Dog or Person Affect Performance of a Working Memory Task?

ABSTRACT

Working memory (WM) plays a critical role in the execution of a wide variety of cognitive tasks and predicts academic success. This study was designed to compare the impact of the presence of a dog or a person, and physical contact with them, on the performance of a WM task. It also exam- ined whether the impact differed for two dogs, and whether these factors im- pacted arousal during the WM task. College students (n=31, aged 18–23 years) performed a WM task in five counterbalanced conditions; dog-touch, dog-no-touch, person-touch, person-no-touch, and alone. Participants were randomly assigned to one of two dogs; Miniature Poodle (n=16) or Border Collie (n=15). The WM task involved replicating increasingly complicated se- quences of colored lights by touching them on an iPad^®. Linear mixed model analyses revealed there was a significant interaction between collaborator and touch (p=0.05); best WM scores occurred without touch with either the per- son or the dog present, and worst WM scores occurred when the participant was touching a dog. Analyzing WM test during the dog conditions, touch (p=0.027) and dog breed (p=0.042) contributed independently to it; task completion was worse when the poodle was present and better without touch. Physiological measures [heart rate (HR) and heart rate variability] during the ex- periment indicated that the WM task was physiologically arousing (p<0.001) compared with listening, and HR was higher when touching a person than a dog during the task (p<0.046). These results are consistent with facilitation of performance by the presence of an observer. If there is a beneficial effect on cognition from a dog, physical contact with the dog might not be a necessary component. Aspects of the dog (e.g., breed) are also likely factors in WM task performance. This study highlights the importance of situational characteristics in studies evaluating the impact of companion animals.     

A nitrogen fertilization field study of carbon-13 and nitrogen-15 transfers in ectomycorrhizas of Pinus sabiniana

Ectomycorrhizal (EM) fungi form relationships with higher plants; plants transfer C to fungi, and fungi transfer nutrients to their host. While evidence indicates that this interaction is largely mutualistic, less is known about how nutrient supply and EM associates may alter C and nutrient exchanges, especially in intact plant-soil-microbe systems in the field. In a dual-labeling experiment with N fertilization, we used C and N stable isotopes to examine in situ transfers in EM pine trees in a Pinus sabiniana woodland in northern California. We added ¹⁵NH₄SO₂ and ¹³CO₂ to track ¹³C transfer from pine needles to EM roots and ¹⁵N transfer from soil to EM roots and pine needles. Transfers of ¹³C and ¹⁵N differed with EM morphotype and with N fertilization. The brown morphotype received the least C per unit of N transferred (5:1); in contrast red and gold morphotypes gained more C and transferred less N (17:1 and 25:1, respectively). N fertilization increased N retention by ectomycorrhizas (EMs) but did not increase N transfer from EMs to pine needles. Therefore N fertilization positively affected both nutrient and C gains by EMs, increasing net C flows and N retention in EMs. Our work on intact and native trees/EM associations thereby extends earlier conclusions based on pot studies with young plants and culturable EM fungi; our results support the concept that EM-host relationships depend on species-level differences as well as responses to soil resources such as N.     

Salt leaching leads to drier soils in disturbed semiarid woodlands of central Argentina

Disturbances in semiarid environments have revealed a strong connection between water, salt and vegetation dynamics highlighting how the alteration of water fluxes can drive salt redistribution process and long-term environmental degradation. Here, we explore to what extent the reciprocal effect, that of salt redistribution on water fluxes, may play a role in dictating environmental changes following disturbance in dry woodlands. We assessed salt and water dynamics comparing soil-solution electrical conductivity, chloride concentration, soil water content (SWC) and soil matric and osmotic water potential (Ψ_m, Ψ_os) between disturbed and undisturbed areas. A large pool of salts and chlorides present in undisturbed areas was absent in disturbed plots, suggesting deep leaching. Unexpectedly, this was associated with slight but consistently lower SWC in disturbed versus undisturbed situations during two growing seasons. The apparent paradox of increased leaching but diminishing SWC after disturbance can be explained by the effect of native salt lowering Ψ_os enough to prevent full soil drying. Under disturbed conditions, the onset of deep drainage and salt leaching would raise Ψ_os allowing a decline of Ψ_m and SWC. Soil water storage seems to be modulated by the presence (under natural conditions) and partial leaching (following selective shrub disturbance) of large salt pools. This counterintuitive effect of disturbances may be important in semiarid regions where deep soil salt accumulation is a common feature. Our results highlight the importance of water–salt–vegetation coupling for the understanding and management of these systems.     

Morphometric and molecular variation in concert: taxonomy and genetics of the reticulate Pyrenean and Iberian alpine spiny fescues (Festuca eskia complex,Poaceae)

The Iberian mountain spiny fescues are a reticulate group of five diploid grass taxa consisting of three parental species and two putative hybrids: F. × souliei (F. eskia × F. quadriflora) and F. × picoeuropeana (F. eskia × F. gautieri). Phenotypic and molecular studies were conducted with the aim of determining the taxonomic boundaries and genetic relationships of the five taxa and disentangling the origins of the two hybrids. Statistical analyses of 31 selected phenotypic traits were conducted on individuals from 159 populations and on nine type specimens. Molecular analyses of random amplified polymorphic DNA (RAPD) markers were performed on 29 populations. The phenotypic analyses detected significant differences between the five taxa and demonstrated the overall intermediacy of the F. × picoeuropeana and F. × souliei between their respective parents. The RAPD analysis corroborated the genetic differentiation of F. eskia, F. gautieri and F. quadriflora and the intermediate nature of the two hybrids; however, they also detected genetic variation within F. × picoeuropeana. These results suggest distinct origins for F. × picoeuropeana in the Cantabrian and Pyrenean mountains, with the sporadic Pyrenean populations having potentially resulted from recent hybridizations and the stabilized Cantabrian ones from older events followed by potential displacements of the parents. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173 , 676–706.     

Human Dietary Exposure to Uranium in Catalonia, Spain

The purpose of this study was to determine the daily dietary intake of uranium (U) by the general population of Catalonia, Spain. Uranium concentrations were measured in foods widely consumed by the population living in that autonomous community. Food samples were randomly acquired in 12 representative cities of Catalonia. The dietary intake of U was estimated for various age–gender groups: children, adolescents, adults, and seniors. Fish and seafood was the food group showing the highest U concentrations (0.090 μg/g of fresh weight (fw)), followed by dairy products (0.044 μg/g fw). In contrast, the lowest U levels were found in oils and fats (0.003 μg/g fw), while in tubers and milk, U was not detected in any sample. The estimated dietary intake of U for a standard male adult of 70 kg body weight living in Catalonia was 15.48 μg/day. According to the age/gender of the population, the highest dietary intake of U corresponded to children (20.32 μg/day), while senior females was the subgroup with the lowest U intake (10.04 μg/day). Based on the tolerable daily intake established for U, the current dietary intake of this metal by the general population of Catalonia should not mean health risks for any of the different age/gender groups of consumers.     

Microtubules that form the stationary lattice of muscle fibers are dynamic and nucleated at Golgi elements

Skeletal muscle microtubules (MTs) form a nonclassic grid-like network, which has so far been documented in static images only. We have now observed and analyzed dynamics of GFP constructs of MT and Golgi markers in single live fibers and in the whole mouse muscle in vivo. Using confocal, intravital, and superresolution microscopy, we find that muscle MTs are dynamic, growing at the typical speed of ∼9 µm/min, and forming small bundles that build a durable network. We also show that static Golgi elements, associated with the MT-organizing center proteins γ-tubulin and pericentrin, are major sites of muscle MT nucleation, in addition to the previously identified sites (i.e., nuclear membranes). These data give us a framework for understanding how muscle MTs organize and how they contribute to the pathology of muscle diseases such as Duchenne muscular dystrophy.