Probing for Binding Regions of the FtsZ Protein Surface through Site-Directed Insertions: Discovery of Fully Functional FtsZ-Fluorescent Proteins

FtsZ, a bacterial tubulin homologue, is a cytoskeletal protein that assembles into protofilaments that are one subunit thick. These protofilaments assemble further to form a “Z ring” at the center of prokaryotic cells. The Z ring generates a constriction force on the inner membrane and also serves as a scaffold to recruit cell wall remodeling proteins for complete cell division in vivo. One model of the Z ring proposes that protofilaments associate via lateral bonds to form ribbons; however, lateral bonds are still only hypothetical. To explore potential lateral bonding sites, we probed the surface of Escherichia coli FtsZ by inserting either small peptides or whole fluorescent proteins (FPs). Among the four lateral surfaces on FtsZ protofilaments, we obtained inserts on the front and back surfaces that were functional for cell division. We concluded that these faces are not sites of essential interactions. Inserts at two sites, G124 and R174, located on the left and right surfaces, completely blocked function, and these sites were identified as possible sites for essential lateral interactions. However, the insert at R174 did not interfere with association of protofilaments into sheets and bundles in vitro. Another goal was to find a location within FtsZ that supported insertion of FP reporter proteins while allowing the FtsZ-FPs to function as the sole source of FtsZ. We discovered one internal site, G55-Q56, where several different FPs could be inserted without impairing function. These FtsZ-FPs may provide advances for imaging Z-ring structure by superresolution techniques. IMPORTANCE One model for the Z-ring structure proposes that protofilaments are assembled into ribbons by lateral bonds between FtsZ subunits. Our study excluded the involvement of the front and back faces of the protofilament in essential interactions in vivo but pointed to two potential lateral bond sites, on the right and left sides. We also identified an FtsZ loop where various fluorescent proteins could be inserted without blocking function; these FtsZ-FPs functioned as the sole source of FtsZ. This advance provides improved tools for all fluorescence imaging of the Z ring and may be especially important for superresolution imaging

Nonstandard Work in Developed Economies: Causes and Consequences

This book reveals the considerable variation in the levels of growth in a broad set of nonstandard work arrangements while presenting a comprehensive view of how, as a result, the nature of the employment relationship is changing within and among countries. The international roster of economists, sociologists, and labor law experts who contributed draw on cross-country variations in economic conditions and institutional characteristics to explain why some arrangements have grown faster in some countries than in others and what this means for workers. By considering a broad array of nonstandard work arrangements in a number of economies, the authors provide a richer understanding than if the focus had been limited to a single country of one or a short-list of employment arrangements.https://research.upjohn.org/up_press/1174/thumbnail.jp

Construction of testers for reversion assay.

Construction of testers for reversion assay

Are both agricultural intensification and farmland abandonment threats to biodiversity? A test with bird communities in paddy-dominated landscapes

Land-use changes, including agricultural intensification and farmland abandonment, influence biodiversity in agricultural landscapes. However, few studies have focused on how the two major land-use changes affect different types of species at landscape scales. This study examined the relationships between the richness and abundance of five bird groups (agricultural wetland species, agricultural land species, grassland species, edge species, and woodland species) as well as the total species richness and abundance, and intensification or abandonment in 28 square, 100-ha grid cells in paddy-dominated landscapes in the Tone River basin of central Japan. Rice-field intensification and abandonment were not completely segregated spatially: intensification occurred in both plain and hilly areas surrounded by forests, while abandonment tended to occur in hilly areas. The effects of intensification and abandonment differed among species groups and between seasons. The richness or abundance of agricultural wetland species in summer were negatively associated with both intensification and abandonment. While the abundance of agricultural land species in winter and grassland species in both seasons were positively associated with intensification and abandonment, respectively. The total species richness and abundance did not show clear association with intensification and abandonment due to a variety of responses of the five bird groups. Based on prefectural Red Data Books, agricultural wetland species, followed by grassland species, were more threatened than other three groups in both summer and winter. This study found that (1) the diversity of habitats (including consolidated and abandoned farmlands) provides buffer areas for the different bird groups on different times of the year and (2) agricultural wetland species that use flooded rice fields in summer, such as egrets and shorebirds, are particularly threatened by both intensification and abandonment.We thank Yoshinori Tokuoka, Susumu Yamada, Eun-Young Kim, and Shori Yamamoto for providing land-use data. We also appreciate two anonymous reviewers for their constructive comments and suggestions. This study was conducted as part of the research project “Developing management techniques for agricultural and aquatic ecosystems in river basins in pursuit of coexistence with nature,” funded by the Agriculture, Forestry and Fisheries Research Council of Japan. N.K. and T.O. were also financially supported by JSPS (Japan Society for the Promotion of Science) KAKENHI Grant Numbers 25830154 and 24710038, respectively. T.A. was supported by the European Commission’s Marie Curie International Incoming Fellowship Programme (PIIFGA-2011-303221).This is the author accepted manuscript. The final version is available from Elsevier via http://dx.doi.org/10.1016/j.agee.2015.08.01

Intraguild predation by Hippodamia variegata (Col.: Coccinellidae) on the parasitoid wasp, Lysiphlebus fabarum (Hym.: Braconidae)

Intraguild predation is defined as killing and eating another predator species as potential competitors that depends upon the same or limited resources. In this study, intraguild predation between the ladybird Hippodamia variegata Goeze and the parasitoid wasp Lysiphlebus fabarum (Marshall) was investigated in association with cotton aphid, Aphis gossypii, on cucumber plants. Host preference for healthy or previously parasitized aphids in third instar larvae and adult females of H. variegata was studied in a choice access situation as well as in a Y-tube olfactometer bioassay. We cut an antenna of each experimental aphid to reliably distinguish  the parasitized aphids from healthy ones. All experiments were performed at 22±1°C, 60%–70% RH, and a 16:8 (L:D) photoperiod. Based on the pre-test results, cutting the antenna of aphids, did not significantly affect the host preference of adults or third instar larvae of ladybirds. In a choice access test, female ladybirds significantly preferred healthy aphids to previously parasitized ones (24 and 72 hours: t =2.15, df =1,18, P =0.04 and t =6.35, df =1,18, P <0.001, respectively), but third instar larvae behaved differently  (t =0.58, df =1,18, P =0.56). According to olfactometer test results, female ladybirds preferred neither healthy aphids nor parasitized ones (χ2 =1.6, df =1,19, P =0.21). It is concluded that preference for healthy aphids over parasitized ones by adult female of H. variegata  facilitates the simultaneous application of the both biocontrol agents, H. variegata and L. fabarum

The Patterns of High-Level Magnetic Activity Occurring on the Surface of V1285 Aql: The OPEA Model of Flares and DFT Models of Stellar Spots

Statistically analyzing Johnson UBVR observations of V1285 Aql during the three observing seasons, both activity level and behavior of the star are discussed in respect to obtained results. We also discuss the out-of-flare variation due to rotational modulation. Eighty-three flares were detected in the U-band observations of season 2006 . First, depending on statistical analyses using the independent samples t-test, the flares were divided into two classes as the fast and the slow flares. According to the results of the test, there is a difference of about 73 s between the flare-equivalent durations of slow and fast flares. The difference should be the difference mentioned in the theoretical models. Second, using the one-phase exponential association function, the distribution of the flare-equivalent durations versus the flare total durations was modeled. Analyzing the model, some parameters such as plateau, half-life values, mean average of the flare-equivalent durations, maximum flare rise, and total duration times are derived. The plateau value, which is an indicator of the saturation level of white-light flares, was derived as 2.421{\pm}0.058 s in this model, while half-life is computed as 201 s. Analyses showed that observed maximum value of flare total duration is 4641 s, while observed maximum flare rise time is 1817 s. According to these results, although computed energies of the flares occurring on the surface of V1285 Aql are generally lower than those of other stars, the length of its flaring loop can be higher than those of more active stars.Comment: 44 pages, 10 figures, 5 tables, 2011PASP..123..659

Neural Advantages of Older Musicians Involve the Cerebellum: Implications for Healthy Aging Through Lifelong Musical Instrument Training

This study compared 30 older musicians and 30 age-matched non-musicians to investigate the association between lifelong musical instrument training and age-related cognitive decline and brain atrophy (musicians: mean age 70.8 years, musical experience 52.7 years; non-musicians: mean age 71.4 years, no or less than 3 years of musical experience). Although previous research has demonstrated that young musicians have larger gray matter volume (GMV) in the auditory-motor cortices and cerebellum than non-musicians, little is known about older musicians. Music imagery in young musicians is also known to share a neural underpinning [the supramarginal gyrus (SMG) and cerebellum] with music performance. Thus, we hypothesized that older musicians would show superiority to non-musicians in some of the abovementioned brain regions. Behavioral performance, GMV, and brain activity, including functional connectivity (FC) during melodic working memory (MWM) tasks, were evaluated in both groups. Behaviorally, musicians exhibited a much higher tapping speed than non-musicians, and tapping speed was correlated with executive function in musicians. Structural analyses revealed larger GMVs in both sides of the cerebellum of musicians, and importantly, this was maintained until very old age. Task-related FC analyses revealed that musicians possessed greater cerebellar-hippocampal FC, which was correlated with tapping speed. Furthermore, musicians showed higher activation in the SMG during MWM tasks; this was correlated with earlier commencement of instrumental training. These results indicate advantages or heightened coupling in brain regions associated with music performance and imagery in musicians. We suggest that lifelong instrumental training highly predicts the structural maintenance of the cerebellum and related cognitive maintenance in old age

In-situ infrared spectroscopy applied to the study of the electrocatalytic reduction of CO2 : Theory, practice and challenges

Acknowledgements RK and WAS thank the European Research Council (ERC) for the financial support under the European Union's Horizon 2020 research and innovation programme (grant agreement no. 759743 – WUTANG). AC acknowledges the support of the Leverhulme Trust (RPG-2015-040) and OA thanks the University of Aberdeen for an Elphinstone Fellowship.Peer reviewedPostprin