Differential sensitivity to ethidium bromide of replicative DNA synthesis and bleomycin-induced unscheduled DNA synthesis in permeable mouse sarcoma cells1

Summary Replicative DNA synthesis in permeable mouse sarcoma cells was more sensitive to ethidium bromide (EtBr) than bleomycin-induced unscheduled DNA synthesis (UDS). A similar difference in sensitivity to EtBr was observed between DNA polymerases and . The difference in sensitivity to EtBr of replicative DNA synthesis and UDS in the present system seems to reflect mainly the sensitivity difference between DNA polymerases and .Acknowledgments. The authors wish to thank Nippon Kayaku Co. (Tokyo, Japan) for providing copper-free bleomycin A₂. This research was supported in part by a grant from the Japan Ministry of Education, Science and Culture.     

基因重组质粒稳定性与苯丙氨酸发酵的研究

本研究应用具有苯丙氨酸生产基因 pheA~(FR)、aroF~(FR)、CI_(857) 的质粒 pSY130～14和质粒 pS Y16,重组构建了具有苯丙氨酸生产基因系统的新质粒 pSY200-14。然后,使其转化到大肠菌 AT2471中,育成了基因重组菌株 AT2471/pSY200-14。试验表明,新菌株质粒稳定性比原菌株有较大的提高。在2. 5升通气搅拌罐进行苯丙氨酸发酵试验,在搅拌转速850rpm、通气速率1. 0VVM,38. 5℃和 pH7. 0的条件下,发酵48小时苯丙氨酸生成量达14. 2g/L,比原菌株 AT2471/pSY130-14增产11. 8%。     

In vitro development ofXenopus skin glands producing 5-hydroxytryptamine and caerulein

The granular glands of amphibian skin synthesize and store a large amount of bioactive amines and peptides which are structurally similar to mammalian brain-gut peptides. To investigate the development of peptide- and amine-producing cells in the granular glands, pieces of dorsal skin taken at various stages fromXenopus laevis tadpoles were cultured, and the contents of caerulein and 5-hydroxytryptamine (5-HT) were measured. When pieces of skin from tadpoles at stages 57 to 60 (Nieuwkoop and Faber stages) were cultured in a medium containing 10% fetal calf serum (FCS medium) or one containing FCS treated with charcoal (chFCS medium), the caerulein and 5-HT levels were increased for the six days of the incubation period. The caerulein content was lower in the chFCS medium than in the FCS medium. Addition of thyroxine to the chFCS medium had no significant effect on the caerulein content. These results show that the caerulein-and 5-HT-producing cells of the granular glands can develop in a culture system with FCS- or chFCS-containing media, and suggest that FCS contains substances which are absorbed by charcoal and stimulate development of the amine- and peptideproducing cells of the glands. In a preliminary search for correlation between caerulein and 5-HT synthesis, addition of 5-hydroxytryptophan (5-HTP), a precursor to 5-HT, to the FCS medium increased 5-HT content and, conversely, caused significant decrease in caerulein content, suggesting that accumulation of caerulein in the granular glands is influenced by the amount of 5-HT synthesis. These studies indicate that this culture system is a useful model for investigating the development of peptide- and amine-producing cells.     

Spontaneous rhythmic contractions in isolated human coronary arteries

Summary Isolated post mortem human coronary arteries developed rhythmic contractions in physiological saline solution without being exposed to vasoactive agents.This study was supported by a Scientific Research Fund (No. 56480181) from the Ministry of Education, Japan.     

Structural organization of the rat thy-1 gene

Thy-1 is a differentiation marker expressed predominantly on thymocytes, T cells and brain tissue. Its presence on murine peripheral T cells but not B cells has long been used to distinguish between these two populations of lymphocytes. Although analogues of Thy-1 have been described in several mammalian species, its tissue distribution in different species varies widely, precluding its use as T-cell-specific marker. The Thy-1 molecule is a cell-surface glycoprotein of relative molecular mass 18,000, one-third of which represents carbohydrate; the protein moieties of the rat and murine Thy-1 molecules have been sequenced and found to consist of 111 and 112 amino acids, respectively. An unusual aspect of Thy-1 is the apparent absence of a hydrophobic segment comparable to that observed in other membrane glycoproteins which would allow integration of Thy-1 within the membrane lipid bilayer. This has prompted speculation that Thy-1 is anchored to the cell surface by some other hydrophobic component such as glycolipid. Here we report the structure of thy-1 complementary DNA and genomic clones and describe the exon-intron organization of the gene. More importantly, our data indicate that Thy-1 is initially synthesized as a molecule of 142 amino acids, 31 amino acids longer at the carboxyl end than the Thy-1 molecule isolated and characterized by Campbell et al. An extremely hydrophobic region of 20 amino acids lies within this 31-amino acid stretch and may represent the transmembrane segment responsible for anchoring Thy-1 to the cell membrane.     

Paradoxical effect of 1-β-D-arabinofuranosyl cytosine triphosphate on bleomycin-induced unscheduled DNA synthesis in permeable sarcoma cells

Summary 1--D-Arabinofuranosyl cytosine-5-triphosphate (araCTP), an inhibitor of DNA synthesis, paradoxically enhanced unscheduled DNA synthesis (USD) induced by bleomycin in permeable mouse sarcoma cells. A greater enhancing effect of araCTP on bleomycin-induced USD was observed with lower concentrations of dCTP in the assay mixture. USD measured without bleomycin in nuclei isolated from mouse sarcoma cells was not enhanced, but inhibited by araCTP.Acknowledgments. The authors wish to thank Nippon Kayaku Co. (Tokyo, Japan) for providing copper-free bleomycin A₂. This research was supported in part by a grant from the Japan Ministry of Education, Science and Culture.     

Terrestrial nitrogen and noble gases in lunar soils

The nitrogen in lunar soils is correlated to the surface and therefore clearly implanted from outside. The straightforward interpretation is that the nitrogen is implanted by the solar wind, but this explanation has difficulties accounting for both the abundance of nitrogen and a variation of the order of 30 per cent in the 15N/14N ratio. Here we propose that most of the nitrogen and some of the other volatile elements in lunar soils may actually have come from the Earth's atmosphere rather than the solar wind. We infer that this hypothesis is quantitatively reasonable if the escape of atmospheric gases, and implantation into lunar soil grains, occurred at a time when the Earth had essentially no geomagnetic field. Thus, evidence preserved in lunar soils might be useful in constraining when the geomagnetic field first appeared. This hypothesis could be tested by examination of lunar farside soils, which should lack the terrestrial component.     

Identification of CRE1 as a cytokinin receptor from Arabidopsis

Cytokinins are a class of plant hormones that are central to the regulation of cell division and differentiation in plants. It has been proposed that they are detected by a two-component system, because overexpression of the histidine kinase gene CKI1 induces typical cytokinin responses and genes for a set of response regulators of two-component systems can be induced by cytokinins. Two-component systems use a histidine kinase as an environmental sensor and rely on a phosphorelay for signal transduction. They are common in microorganisms, and are also emerging as important signal detection routes in plants. Here we report the identification of a cytokinin receptor. We identified Arabidopsis cre1 (cytokinin response 1) mutants, which exhibited reduced responses to cytokinins. The mutated gene CRE1 encodes a histidine kinase. CRE1 expression conferred a cytokinin-dependent growth phenotype on a yeast mutant that lacked the endogenous histidine kinase SLN1 (ref. 10), providing direct evidence that CRE1 is a cytokinin receptor. We also provide evidence that cytokinins can activate CRE1 to initiate phosphorelay signalling.