Characterization of dicyclohexylcarbodiimide binding sites in beef-heart mitochondria.

Binding studies with [¹⁴C]-dicyclohexylcarbodiimide showed the presence of binding sites in the beef-heart mitochondrial membrane at a concentration of 1.8 nmol/mg protein (1.4 sites per cytochrome a+a₃). Saturation of these sites correlated with the inhibition of the ATPase activity. The maximum binding capacity could be related with the amount of F₁-ATPase in mitochondria from various tissues.     

Effects of alkylating agents on lymphocytes from controls and from patients with Fanconi's anemia

Summary The frequency of sister chromatid exchanges (SCE) and chromosome aberrations and the dynamics of cell division in peripheral blood lymphocytes of four patients with Fanconi's anemia were studied after in vitro exposure to alkylating agents TEPA and mitomycin.SCE frequency was significantly increased even after very low doses of mutagens, while chromosome aberrations were significantly increased only after high doses (0.160 g/ml mitomycin and 10^-5 M TEPA). The responses of Fanconi's anemia cells and control cells did not differ significantly. The increased frequency of both SCE and chromosome aberrations was accompanied by gradual delay of cell division, which was most conspicuous in cells from patients with Fanconi's anemia.     

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Genetic and physiological variability among cultured cells and regenerated plants of Nicotiana tabacum

Summary Tobacco cell lines selected for resistance to picloram (4-amino-3,5,6-trichloropicolinic acid) and plants regenerated from these cell lines manifest several traits not shown by the parental strains. Genetics analyses of the regenerated plants have permitted the sources of this variability to be identified.Tricotyledenous seedlings appeared at a much higher frequency among the progeny of a heterozygous mutant plant (PmR1/+) regenerated from culture than they did among progeny of normal regenerated plants. In crosses with the regenerated heterozygous mutant plant and with homozygous progeny of this plant (PmR1/PmR1) the frequency of tricotyly was influenced more by the generation than by the genotype of the parent plant. Therefore, it is concluded that tricotyly is a physiological response to passage through cell culture.More than half of the picloram-resistant cell lines isolated were also resistant to hydroxyurea. Segregation of these two resistances was analyzed in progeny of crosses with regenerated plants. In all cases hydroxyurea-resistance was genetically stable and inherited as a single dominant nuclear mutation (designated HuR). In crosses with plants PmR1/+ and PmR7/+ the HuR and PmR mutations assorted independently. In contrast, the HuR mutation recovered from plant PmR6/+ was linked to the PmR6 mutation.     

Transgelin Silencing Induces Different Processes in Different Breast Cancer Cell Lines

Transgelin is a protein reported to be a marker of several cancers. However, previous studies have shown both up‐ and down‐regulation of transgelin in tumors when compared with non‐tumor tissues and the mechanisms whereby transgelin may affect the development of cancer remain largely unknown. Transgelin is especially abundant in smooth muscle cells and is associated with actin stress fibers. These contractile structures participate in cell motility, adhesion, and the maintenance of cell morphology. Here, the role of transgelin in breast cancer is focused on. Initially, the effects of transgelin on cell migration of the breast cancer cell lines, BT 549 and PMC 42, is studied. Interestingly, transgelin silencing increased the migration of PMC 42 cells, but decreased the migration of BT 549 cells. To clarify these contradictory results, the changes in protein abundances after transgelin silencing in these two cell lines are analyzed using quantitative proteomics. The results confirmed the role of transgelin in the migration of BT 549 cells and suggest the involvement of transgelin in apoptosis and small molecule biochemistry in PMC 42 cells. The context‐dependent function of transgelin reflects the different molecular backgrounds of these cell lines, which differ in karyotypes, mutation statuses, and proteome profiles.     

Transmembrane adaptor protein WBP1L regulates CXCR4 signalling and murine haematopoiesis

WW domain binding protein 1‐like (WBP1L), also known as outcome predictor of acute leukaemia 1 (OPAL1), is a transmembrane adaptor protein, expression of which correlates with ETV6‐RUNX1 (t(12;21)(p13;q22)) translocation and favourable prognosis in childhood leukaemia. It has a broad expression pattern in haematopoietic and in non‐haematopoietic cells. However, its physiological function has been unknown. Here, we show that WBP1L negatively regulates signalling through a critical chemokine receptor CXCR4 in multiple leucocyte subsets and cell lines. We also show that WBP1L interacts with NEDD4‐family ubiquitin ligases and regulates CXCR4 ubiquitination and expression. Moreover, analysis of Wbp1l‐deficient mice revealed alterations in B cell development and enhanced efficiency of bone marrow cell transplantation. Collectively, our data show that WBP1L is a novel regulator of CXCR4 signalling and haematopoiesis.     

Phenology and polyploidy in annual Brachypodium species (Poaceae) along the aridity gradient in Israel

Local adaptation of plants along environmental gradients provides strong evidence for clinal evolution mediated by natural selection. Plants have developed diverse strategies to mitigate stress, for example, drought escape is a phenological strategy to avoid drought stress, while polyploidy was proposed as a genomic adaptation to stress. Polyploidy as an adaptation to aridity (an environmental parameter integrating temperature and precipitation) was previously documented in annual Brachypodium spp. (Poaceae) in the Western Mediterranean. Here, we examined whether polyploidy or phenology are associated with aridity in annual Brachypodium spp. along the aridity gradient in the Eastern Mediterranean. Using flow cytometry, we determined ploidy levels of plants from natural populations along the Israeli gradient, spanning ∼424 km from mesic Mediterranean to extreme desert climates. In a common garden we recorded time of seedling emergence, flowering and senescence. We tested whether the proportion of allotetraploids in the populations and phenological traits were associated with aridity. Contrary to a previous study in the Western Mediterranean, we found no effect of aridity on the proportion of allotetraploids and diploids within populations. Interestingly, phenology was associated with aridity: time of emergence was later, while flowering and senescence were earlier in desert plants. Our results indicate that in the Eastern Mediterranean, adaptation of Brachypodium to aridity is mediated mainly by phenology, rather than ploidy level. Therefore, we suggest that genome duplication is not the main driver of adaptation to environmental stress; rather, phenological change as a drought escape mechanism may be the major adaptation.     

Leaving uncut refuges during meadow harvesting increases the functional diversity of Orthoptera

In the past, insect species richness was high in Central European seminatural grasslands, which were characterized by low‐intensity land use. Currently, however, the hay in most of these grasslands is mechanically harvested, which negatively impacts insect biodiversity. One way to reduce this negative effect is to leave unmown patches as refuges. In the current research we evaluated the short‐term effects of leaving an unmown patch on the taxonomic and functional diversity of the Orthoptera assemblage in a meadow. We found that orthopteran species richness and abundance were significantly reduced by mowing, whether or not a patch was left uncut. In contrast, functional evenness, indicating distribution of species abundances in a niche space, was reduced by mowing only if the plot lacked an uncut refuge. Functional richness, indicating the amount of niche space occupied by species, was elevated if the plot had an uncut refuge. Larger species were negatively affected by mowing, while habitat specialists, mobile species and soil‐ovipositing species benefitted from it. We infer that the presence of an uncut patch increased the diversity of habitats available to orthopterans and maintained even distribution of species among niche space. In summary, leaving an unmown refuge in grasslands could increase the functional diversity of orthopterans, even if it does not preserve taxonomic diversity.