Factor-free and one-factor-promoted poly(U,C)-dependent synthesis of polypeptides in cell-free systems from Escherichia coli

α-Tropomyosin from rat cardiac muscle was shown by two-dimensional gel electrophoresis to become phosphorylated when tissue slices were incubated in Eagle's medium supplemented with ³²P_i. In the adult rat and mouse heart the level of phosphorylation was ～30%, but the level was much higher in the foetal heart (60–70%). A similar developmental trend was observed in skeletal muscle from the rat and mouse, where phosphorylated forms of both α- and β-tropomyosins were observed. When rat cardiac cells were grown in tissue culture in the presence of ³²P_i, radioactivity was incorporated into the region of the gel containing tropomyosin.     

Toxic ligand conjugates as tools in the study of receptor-ligand interactions

We have constructed hybrid proteins in which the toxic A chains of ricin or diptheria toxin have been linked to either asialofetuin, fetuin, or epidermal growth factor (EGF). Both ASF-RTA and ASF-DTA are potent toxins on cultured rat hepatocytes, cells that display the asialoglycoprotein receptor. Toxicity of these two compounds is restricted to hepatocytes and can be blocked by asialoglycoproteins but not the native glycoproteins or asialoagalactoglycoprotein derivatives, indicating that the toxicity of the conjugates is mediated by the hepatic asialoglycoprotein receptor. The EGF-RTA conjugate is an extremely potent toxin on cells that can bind the hormone, but is only poorly effective on cells that are unable to bind EGF. The EGF-DTA conjugate, in contrast, is unable to kill 3T3 cells and is at least two orders of magnitude less effective than EGF-RTA on A431 cells, a cell line with 1-2 X 10(6) EGF receptors per cell. However, when EGF-RTA and EGF-DTA were tested on primary liver hepatocyte cultures, which were susceptible to both ASF-RTA and ASF-DTA, both EGF conjugates were potent toxins. Sensitivity of the hepatocyte cultures to ricin toxicity increases slightly during a 52-hr culture period. In contrast, sensitivity to EGF-RTA and ASF-RTA decline dramatically during this period. Receptors for both ligands remain plentiful on the cell surface during this time.     

Estimation of phenological stages and physiological states of grasslands from remote sensing data

Two radiometers measuring reflectance factors have been used at a height of 1.50 m above some grasslands in C. France. The results show that both spectral data are sensitive to the photosynthetic activity of the grassland. Measurements made in April, May, June and July show that grasslands have quite different spectral behaviour according to soil conditions or to grazing level. Grasslands on dry or wet soils may be separated from those of normal soils for which overgrazing and trampling affecting the growth of species, are shown by the different spectral values. Such on the ground remote sensing measurements may then be proposed for evaluating the range of growth and development of different grasslands.Convention C.N.E.S./I.N.A.P-G. ref.: 80/CNES/24 I thank CNES and IGN for the facilities given to assume this experiment. Many thanks to Lynn Erselius, who revised the text. Mr. Lecordix, a student from the Ecole Nationale des Sciences Géographiques and Mr. Besnard, a student from the Orsay University (Plant Biology) helped in the collection and handling of spectral resp, floristical data.     

Investigations of 'transfer factor' activity in immunity to Ostertagia circumcincta and Trichostrongylus colubriformis infections in sheep.

Immunity was successfully transferred by ‘Transfer Factor’ prepared from leucocytes of two adult Scottish Blackface donor rams infected with O. circumcincta and T. colubriformis to 4-month-old susceptible Fin X Dorset lambs. The immunity was expressed by a significantly reduced faecal egg count and worm burden compared to challenged, untreated controls. The immunity was comparable to that produced in another group of lambs given an initial infection prior to challenge with both parasites.     

Stimulation of human vascular endothelial cell growth by a platelet-derived growth factor and thrombin

Repair of a vascular wound is mediated by migration and subsequent replication of the endothelial cells that form the inner lining of blood vessels. We have measured the growth response of human umbilical vein endothelial cells (HuE) to two polypeptides that are transiently produced in high concentrations at the site of a wound; the platelet-derived growth factor (PDGF) and the protease thrombin. When 10⁴ HuE cells are seeded as a dense island (2-mm diameter) in the center of a 16-mm tissue culture well in medium containing 20% human serum derived from platelet-poor plasma (PDS), no increase in cell number or colony size is observed. With the addition of 0.5 ng/ml partially purified PDGF, colony size increases and the number of cells after 8 days is 4.8 × 10⁴. When human thrombin (1 μg/ml) is added along with the PDGF, the cell number rises to 9.2 × 10⁴. Thrombin alone stimulates no increase in cell number. Although partially purified PDGF stimulates endothelial cells maintained in PDS as well as those maintained in whole blood serum (WBS), pure PDGF is active only when assayed in medium that contains WBS and is supplemented with thrombin. These results suggest the existence of a second class of platelet-derived factors that enable HuE cells to respond to the mitogenic activity of the purified platelet mitogen and thrombin.     

mTORC1-independent translation control in mammalian cells by methionine adenosyltransferase 2A and S-adenosylmethionine

Methionine adenosyltransferase (MAT) catalyzes the synthesis of S-adenosylmethionine (SAM). As the sole methyl-donor for methylation of DNA, RNA, and proteins, SAM levels affect gene expression by changing methylation patterns. Expression of MAT2A, the catalytic subunit of isozyme MAT2, is positively correlated with proliferation of cancer cells; however, how MAT2A promotes cell proliferation is largely unknown. Given that the protein synthesis is induced in proliferating cells and that RNA and protein components of translation machinery are methylated, we tested here whether MAT2 and SAM are coupled with protein synthesis. By measuring ongoing protein translation via puromycin labeling, we revealed that MAT2A depletion or chemical inhibition reduced protein synthesis in HeLa and Hepa1 cells. Furthermore, overexpression of MAT2A enhanced protein synthesis, indicating that SAM is limiting under normal culture conditions. In addition, MAT2 inhibition did not accompany reduction in mechanistic target of rapamycin complex 1 activity but nevertheless reduced polysome formation. Polysome-bound RNA sequencing revealed that MAT2 inhibition decreased translation efficiency of some fraction of mRNAs. MAT2A was also found to interact with the proteins involved in rRNA processing and ribosome biogenesis; depletion or inhibition of MAT2 reduced 18S rRNA processing. Finally, quantitative mass spectrometry revealed that some translation factors were dynamically methylated in response to the activity of MAT2A. These observations suggest that cells possess an mTOR-independent regulatory mechanism that tunes translation in response to the levels of SAM. Such a system may acclimate cells for survival when SAM synthesis is reduced, whereas it may support proliferation when SAM is sufficient.     

转录因子在植物干旱应激中的功能研究进展

转录因子是一种多功能蛋白,在感知应激信号、应答相应应激基因表达及传导应激信号中起着关键作用。干旱是影响植物生长发育的主要非生物胁迫之一。为了适应干旱环境,植物发展了复杂的分子机制,其中转录因子可同时控制多种途径调控干旱应激,是操纵调控和应激响应途径的有力工具。近年来,越来越多的植物转录因子的功能被阐明,了解转录因子在干旱应激的功能,对植物的工程抗旱有重要的实践意义。综述转录因子在植物干旱应激中的功能研究进展,以期为今后转录因子的研究和利用提供理论依据,培育具有较强抗旱能力的植物。